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Kusintha kwa tizilombo tating'onoting'ono kumaphatikizapo kutsutsana pakati pa kusankha kwachilengedwe, komwe kumapangitsa kuti tizilombo toyambitsa matenda tiziyenda bwino, ndi kusuntha kwa majini, zomwe zimapangitsa kuti tizilombo toyambitsa matenda tiwononge majini ndikudziunjikira kusintha koopsa.Apa, kuti timvetsetse momwe kuphatikizikaku kumachitika pamlingo wa macromolecule imodzi, timafotokoza za cryo-EM kapangidwe ka ribosome ya Encephalitozoon cuniculi, chamoyo cha eukaryotic chokhala ndi genome imodzi yaying'ono kwambiri m'chilengedwe.Kuchepetsa kwakukulu kwa rRNA mu E. cuniculi ribosomes kumatsagana ndi kusintha kosaneneka kwa kapangidwe kake, monga kusinthika kwa zolumikizira zolumikizana za rRNA zomwe zidadziwika kale ndi rRNA popanda zotupa.Kuonjezera apo, E. cuniculi ribosome inapulumuka kutayika kwa zidutswa za rRNA ndi mapuloteni popanga luso logwiritsa ntchito mamolekyu ang'onoang'ono monga mafanizidwe apangidwe a zidutswa zowonongeka za rRNA ndi mapuloteni.Ponseponse, tikuwonetsa kuti mamolekyu omwe amaganiziridwa kale kuti amachepetsedwa, amawonongeka, komanso amatha kufooketsa masinthidwe ali ndi njira zingapo zolipirira zomwe zimawapangitsa kukhala achangu ngakhale kuti ma cell achepa kwambiri.
Chifukwa magulu ambiri a tizilombo tating'onoting'ono timakhala ndi zida zapadera zogwiritsira ntchito mabakiteriya awo, nthawi zambiri timayenera kupanga mankhwala osiyanasiyana amagulu osiyanasiyana a tizilombo toyambitsa matenda1,2.Komabe, umboni watsopano ukusonyeza kuti mbali zina za chisinthiko cha tizilombo toyambitsa matenda zimasinthasintha ndipo makamaka zimadziwikiratu, zomwe zimasonyeza kuti pali maziko ochiritsira ochuluka a tizilombo toyambitsa matenda3,4,5,6,7,8,9.
Ntchito yam'mbuyomu yazindikira zachisinthiko chofala mu tizirombo tating'onoting'ono totchedwa kuchepetsa ma genome kapena kuwonongeka kwa genome10,11,12,13.Kafukufuku wamakono akuwonetsa kuti tizilombo toyambitsa matenda tikasiya moyo wawo waufulu ndikukhala tizilombo toyambitsa matenda (kapena endosymbionts), ma genome awo amapita pang'onopang'ono koma modabwitsa metamorphoses pazaka mamiliyoni9,11.Munjira yomwe imadziwika kuti kuwonongeka kwa ma genome, tizilombo tating'onoting'ono timapanga masinthidwe owopsa omwe amasintha majini ambiri ofunikira kukhala ma pseudogenes, zomwe zimatsogolera kutayika pang'onopang'ono kwa majini komanso kugwa kosinthika14,15.Kugwa kumeneku kungathe kuwononga mpaka 95% ya majini mu zamoyo zakale kwambiri za m'kati mwa maselo kuyerekeza ndi zamoyo zaufulu zogwirizana kwambiri.Choncho, kusinthika kwa tizilombo toyambitsa matenda tizilombo toyambitsa matenda ndikumenyana pakati pa mphamvu ziwiri zotsutsana: Kusankhidwa kwa chilengedwe cha Darwin, zomwe zimabweretsa kusintha kwa tizilombo toyambitsa matenda, ndi kugwa kwa genome, kutaya tizilombo toyambitsa matenda.Sizikudziwikabe kuti tizilomboti tinatha bwanji kutuluka m'nkhondo imeneyi n'kusunga mamolekyu ake.
Ngakhale kuti kachitidwe ka kuwonongeka kwa ma genome sikumveka bwino, zikuwoneka kuti zimachitika makamaka chifukwa cha kusuntha kwa chibadwa pafupipafupi.Chifukwa majeremusi amakhala m'magulu ang'onoang'ono, osagonana ndi amuna kapena akazi okhaokha, sangathe kuthetsa masinthidwe owopsa omwe nthawi zina amapezeka pakubwereza kwa DNA.Izi zimabweretsa kudzikundikira kosasinthika kwa masinthidwe owopsa ndikuchepetsa kwa tiziromboti.Chotsatira chake, tizilombo toyambitsa matenda sikuti timangotaya majini omwe salinso ofunikira kuti akhalebe ndi moyo mu chilengedwe.Ndikulephera kwa tizilombo toyambitsa matenda kuti tithe kuthetseratu masinthidwe amtundu uliwonse omwe amachititsa kuti masinthidwe awa achulukane mumtundu wonse wa majeremusi, kuphatikizapo majini ofunika kwambiri.
Zambiri zomwe timamvetsetsa pakuchepetsa ma genome zimangotengera kufananiza kwa ma genome, osayang'ana pang'ono kusintha kwa mamolekyu enieni omwe amagwira ntchito yosamalira m'nyumba ndipo amakhala ngati chandamale chamankhwala.Kafukufuku woyerekeza awonetsa kuti kulemedwa kwa kusintha kwa ma intracellular microbial kumawoneka kuti kumapangitsa kuti mapuloteni ndi ma nucleic acid asokonezeke ndikuphatikizana, kuwapangitsa kukhala odalira kwambiri komanso osamva kutentha19,20,21,22,23.Kuphatikiza apo, majeremusi osiyanasiyana-chisinthiko chodziyimira pawokha nthawi zina cholekanitsidwa ndi zaka 2.5 biliyoni-zidakumananso ndi kutayika kofananira kwa malo owongolera omwe ali mu protein synthesis5,6 ndi njira zokonzera DNA24.Komabe, ndizochepa zomwe zimadziwika za momwe moyo wamkati umakhudzira zinthu zina zonse zama cell macromolecules, kuphatikiza kusintha kwa ma cell ndikuchulukirachulukira kwa masinthidwe owopsa.
Mu ntchito iyi, kuti timvetse bwino kusinthika kwa mapuloteni ndi nucleic zidulo wa okhudza maselo ambiri tizilombo tating'onoting'ono, tinatsimikiza kapangidwe ribosomes wa okhudza maselo ambiri tiziromboti Encephalitozoon cuniculi.E. cuniculi ndi chamoyo chofanana ndi bowa cha gulu la parasitic microsporidia omwe ali ndi ma genome ang'onoang'ono a eukaryotic motero amagwiritsidwa ntchito ngati zamoyo zachitsanzo kuphunzira kuwonongeka kwa ma genome25,26,27,28,29,30.Posachedwapa, mawonekedwe a cryo-EM ribosome adatsimikiziridwa kuti achepetse pang'ono ma genomes a Microsporidia, Paranosema locustae, ndi Vairimorpha necatrix31,32 (~ 3.2 Mb genome).Mapangidwewa akuwonetsa kuti kutayika kwina kwa kukulitsa kwa rRNA kumalipidwa ndi kupangika kwa kulumikizana kwatsopano pakati pa mapuloteni oyandikana nawo a ribosomal kapena kupeza mapuloteni atsopano a msL131,32 ribosomal.Mitundu ya Encephalitozoon (genome ~ 2.5 miliyoni bp), pamodzi ndi wachibale wawo wapamtima wa Ordospora, amawonetsa kuchepa kwakukulu kwa ma genome mu eukaryotes - ali ndi majini opangira mapuloteni ochepera 2000, ndipo akuyembekezeka kuti ma ribosomes awo sangokhala opanda rRNA tizidutswa tating'onoting'ono tomwe timakhala ndi mabakiteriya anayi a RNA omwe ali ndi mabakiteriya anayi mapuloteni a ribosomal chifukwa cha kusowa kwawo kwa ma homologues mu E. cuniculi genome26,27,28.Choncho, tinaganiza kuti E. cuniculi ribosome ikhoza kuwulula njira zomwe sizinali zodziwika kale zosinthira maselo kuti awonongeke.
Kapangidwe kathu ka cryo-EM ndikuyimira kakang'ono kwambiri ka eukaryotic cytoplasmic ribosome yomwe imayenera kudziwika ndipo imapereka chidziwitso cha momwe kuchepa kwa ma genome kumakhudzira kapangidwe kake, kuphatikiza, ndi kusinthika kwa makina amolekyu omwe ali ofunikira mu selo.Tinapeza kuti E. cuniculi ribosome imaphwanya mfundo zambiri zosungidwa za RNA kupukutira ndi kusonkhana kwa ribosome, ndipo tinapeza mapuloteni atsopano, omwe kale anali osadziwika.Mosayembekezereka, tikuwonetsa kuti ma microsporidia ribosomes adasintha kuthekera komanga mamolekyu ang'onoang'ono, ndikuyerekeza kuti kudumpha kwa rRNA ndi mapuloteni kumayambitsa kusinthika komwe kumatha kupereka mikhalidwe yothandiza pa ribosome.
Kuti timvetse bwino za kusinthika kwa mapuloteni ndi nucleic acid mu zamoyo za intracellular, tinaganiza zopatula E. cuniculi spores ku zikhalidwe za maselo amtundu wa mammalian omwe ali ndi kachilombo kuti ayeretse ma ribosomes awo ndi kudziwa momwe ma ribosomes awa.Ndizovuta kupeza kuchuluka kwa parasitic microsporidia chifukwa microsporidia sangathe kulimidwa muzakudya zopatsa thanzi.M’malo mwake, zimakula ndi kuberekana m’kati mwa selo lokhalamo.Choncho, kuti tipeze E. cuniculi biomass yoyeretsa ribosome, tinayambitsa kachilombo ka mammalian kidney cell line RK13 ndi E. cuniculi spores ndipo tinakulitsa maselo omwe ali ndi kachilomboka kwa milungu ingapo kuti E. cuniculi ikule ndi kuchulukana.Pogwiritsa ntchito cell cell monolayer pafupifupi theka la sikweya mita, tinatha kuyeretsa pafupifupi 300 mg wa Microsporidia spores ndikuwagwiritsa ntchito kupatula ribosomes.Kenako tidasokoneza spores zoyeretsedwa ndi mikanda yagalasi ndikupatula ma ribosomes opanda pake pogwiritsa ntchito stepwise polyethylene glycol fractionation of the lysates.Izi zinatilola kupeza pafupifupi 300 µg ya E. cuniculi ribosomes yaiwisi kuti tifufuze kamangidwe.
Kenako tidasonkhanitsa zithunzi za cryo-EM pogwiritsa ntchito zitsanzo za ribosome ndikukonza zithunzizi pogwiritsa ntchito masks ofanana ndi subunit yayikulu ya ribosomal, mutu waung'ono, ndi gawo laling'ono.Panthawiyi, tinasonkhanitsa zithunzi za 108,000 ribosomal particles ndikujambula zithunzi za cryo-EM ndi chisankho cha 2.7 Å (Supplementary Figures 1-3).Kenako tinagwiritsa ntchito zithunzi za cryoEM kuti tiwonetsere rRNA, mapuloteni a ribosomal, ndi hibernation factor Mdf1 yogwirizana ndi E. cuniculi ribosomes (Mkuyu 1a, b).
Mapangidwe a E. cuniculi ribosome mu zovuta ndi hibernation factor Mdf1 (pdb id 7QEP).b Mapu a hibernation factor Mdf1 yolumikizidwa ndi E. cuniculi ribosome.c Mapu achiwiri akuyerekeza rRNA yopezekanso mu mitundu ya Microsporidian ndi zodziwika bwino za ribosomal.Mapanelo akuwonetsa komwe kuli zidutswa za rRNA (ES) ndi malo omwe ali ndi ribosome, kuphatikiza malo opangira ma decoding (DC), sarcinicin loop (SRL), ndi peptidyl transferase center (PTC).d Kuchuluka kwa ma elekitironi kumagwirizana ndi peptidyl transferase center ya E. cuniculi ribosome imasonyeza kuti malo othandizirawa ali ndi dongosolo lomwelo mu E. cuniculi parasite ndi makamu ake, kuphatikizapo H. sapiens.e, f The lolingana ma elekitironi kachulukidwe wa likulu decoding (e) ndi dongosolo schematic wa decoding center (f) zimasonyeza kuti E. cuniculi ndi zotsalira U1491 m'malo A1491 (E. coli manambala) mu eukaryotes ena ambiri.Kusinthaku kukuwonetsa kuti E. cuniculi akhoza kukhala okhudzidwa ndi maantibayotiki omwe amalunjika pamalowa.
Mosiyana ndi mapangidwe omwe adakhazikitsidwa kale a V. necatrix ndi P. locustae ribosomes (zonse ziwirizi zikuimira banja la microsporidia Nosematidae ndipo ndizofanana kwambiri), 31,32 E. cuniculi ribosomes amakumana ndi njira zambiri za rRNA ndi kugawanika kwa mapuloteni.Kutanthauzira kwina (Zithunzi Zowonjezera 4-6).Mu rRNA, kusintha kwakukulu kunaphatikizapo kutayika kwathunthu kwa 25S rRNA fragment ES12L ndi kuchepa pang'ono kwa h39, h41, ndi H18 helices (Mkuyu 1c, Supplementary Fig. 4).Pakati pa mapuloteni a ribosomal, kusintha kochititsa chidwi kunaphatikizapo kutayika kwathunthu kwa mapuloteni a eS30 ndi kufupikitsa eL8, eL13, eL18, eL22, eL29, eL40, uS3, uS9, uS14, uS17, ndi mapuloteni a eS7 (Zowonjezera Zowonjezera 4, 5).
Motero, kuchepa kwakukulu kwa majeremusi a mitundu ya Encephalotozoon/Ordospora kumaonekera m’kapangidwe kake ka ribosome: E. cuniculi ribosomes amataya kwambiri mapuloteni opezeka mu eukaryotic cytoplasmic ribosomes omwe ali ndi mawonekedwe ake, ndipo alibe ngakhale ma rRNA ndi zidutswa za protein zomwe zimasungidwa m'magawo atatu okha, osati ma eukaryotic cytoplasmic ribosomes.Mapangidwe a E. cuniculi ribosome amapereka chitsanzo choyamba cha maselo a kusintha kumeneku ndikuwonetsa zochitika zachisinthiko zomwe zakhala zimanyalanyazidwa ndi ma genomics ofananitsa ndi maphunziro a intracellular biomolecular structure (Supplementary Fig. 7).Pansipa, tikufotokozera chilichonse mwazochitikazi limodzi ndi zomwe zidachitika komanso momwe zingakhudzire ntchito ya ribosome.
Kenako tidapeza kuti, kuwonjezera pa kudulidwa kwakukulu kwa rRNA, ma E. cuniculi ribosomes ali ndi kusiyana kwa rRNA pa amodzi mwa malo omwe amagwira ntchito.Ngakhale kuti peptidyl transferase center ya E. cuniculi ribosome ili ndi dongosolo lofanana ndi ma eukaryotic ribosomes (mkuyu 1d), malo opangira ma decoding amasiyana chifukwa cha kusintha kwa ma nucleotide 1491 (E. coli nambala, Fig. 1e, f).Kuwunikiraku ndikofunikira chifukwa malo opangira ma eukaryotic ribosomes nthawi zambiri amakhala ndi zotsalira za G1408 ndi A1491 poyerekeza ndi zotsalira zamtundu wa bakiteriya A1408 ndi G1491.Kusiyanasiyana kumeneku kumayambitsa kukhudzika kosiyana kwa mabakiteriya ndi ma eukaryotic ribosomes ku gulu la aminoglycoside la maantibayotiki a ribosomal ndi mamolekyu ena ang'onoang'ono omwe amalunjika pamalo opangirako.Pamalo osindikizira a E. cuniculi ribosome, zotsalira za A1491 zinasinthidwa ndi U1491, zomwe zingathe kupanga mawonekedwe apadera a mamolekyu ang'onoang'ono omwe akulunjika pamalowa.Chofanana cha A14901 chimapezekanso mu microsporidia zina monga P. locustae ndi V. necatrix, zomwe zimasonyeza kuti ndizofala pakati pa mitundu ya microsporidia (mkuyu 1f).
Chifukwa zitsanzo zathu za E. cuniculi ribosome zinali zolekanitsidwa ndi spores zosagwira ntchito za kagayidwe kachakudya, tinayesa mapu a cryo-EM a E. cuniculi pofotokoza m'mbuyomo momwe ribosome imamangiriza pansi pa nkhawa kapena njala.Zinthu za hibernation 31,32,36,37, 38. Tinafananiza dongosolo lomwe linakhazikitsidwa kale la hibernating ribosome ndi mapu a cryo-EM a E. cuniculi ribosome.Pa docking, S. cerevisiae ribosomes ankagwiritsidwa ntchito movutikira ndi hibernation factor Stm138, dzombe ribosomes zovuta ndi Lso232 factor, ndi V. necatrix ribosomes zovuta ndi Mdf1 ndi Mdf231 zinthu.Nthawi yomweyo, tapeza kuchuluka kwa cryo-EM kofanana ndi zina zonse Mdf1.Mofanana ndi Mdf1 yomangiriza ku V. necatrix ribosome, Mdf1 imamangirizanso ku E. cuniculi ribosome, kumene imatchinga malo a E a ribosome, mwinamwake kuthandizira kuti ma ribosomes apezeke pamene tizilombo toyambitsa matenda timakhala tikugwira ntchito poyambitsa thupi (Chithunzi 2).).
Mdf1 imatchinga malo a E a ribosome, omwe amawoneka kuti amathandizira kuti ribosome ayambe kugwira ntchito pamene tizilombo toyambitsa matenda timasiya kugwira ntchito.Mu dongosolo la E. cuniculi ribosome, tinapeza kuti Mdf1 imapanga mgwirizano wosadziwika kale ndi tsinde la L1 ribosome, gawo la ribosome lomwe limathandizira kutulutsidwa kwa deacylated tRNA kuchokera ku ribosome panthawi ya mapuloteni.Kulumikizana kumeneku kumasonyeza kuti Mdf1 imasiyanitsidwa ndi ribosome pogwiritsa ntchito njira yofanana ndi deacetylated tRNA, ndikupereka kufotokozera momwe ribosome imachotsera Mdf1 kuti iyambitsenso mapuloteni.
Komabe, mapangidwe athu adawonetsa kulumikizana kosadziwika pakati pa Mdf1 ndi mwendo wa L1 ribosome (gawo la ribosome lomwe limathandiza kutulutsa deacylated tRNA kuchokera ku ribosome panthawi yopanga mapuloteni).Makamaka, Mdf1 imagwiritsa ntchito kukhudzana komweko monga gawo la chigongono cha molekyulu ya deacylated tRNA (mkuyu 2).Mawonekedwe a mamolekyu omwe sanadziwike kale adawonetsa kuti Mdf1 imasiyanitsidwa ndi ribosome pogwiritsa ntchito njira yofanana ndi deacetylated tRNA, yomwe imafotokoza momwe ribosome imachotsera hibernation factor kuti ayambitsenso kaphatikizidwe ka mapuloteni.
Pomanga chitsanzo cha rRNA, tinapeza kuti E. cuniculi ribosome ili ndi zidutswa za rRNA zopindika, zomwe timazitcha kuti rRNA (mkuyu 3).M'ma ribosomes omwe amakhala m'madera atatu a moyo, rRNA imalowa m'magulu omwe ambiri a rRNA amayambira awiri ndi kupindika wina ndi mzake kapena amagwirizana ndi mapuloteni a ribosomal38,39,40.Komabe, mu E. cuniculi ribosomes, ma rRNA akuwoneka kuti akuphwanya mfundo yopindika imeneyi potembenuza ena mwa ma helice awo kukhala zigawo zosasinthika za rRNA.
Mapangidwe a H18 25S rRNA helix mu S. cerevisiae, V. necatrix, ndi E. cuniculi.Nthawi zambiri, m'ma ribosomes omwe amazungulira magawo atatu amoyo, cholumikizira ichi chimazungulira kukhala RNA helix yomwe ili ndi zotsalira 24 mpaka 34.Mu Microsporidia, mosiyana, cholumikizira ichi cha rRNA chimachepetsedwa pang'onopang'ono kukhala maulalo awiri amtundu umodzi wokhala ndi uridine wokhala ndi zotsalira za 12 zokha.Zambiri mwa zotsalirazi zimakhudzidwa ndi zosungunulira.Chiwerengerochi chikuwonetsa kuti parasitic microsporidia ikuwoneka kuti ikuphwanya mfundo za rRNA kupukutira, pomwe maziko a rRNA nthawi zambiri amaphatikizidwa ndi maziko ena kapena amakhudzidwa ndi rRNA-mapuloteni.Mu microsporidia, zidutswa zina za rRNA zimatenga khola losavomerezeka, momwe helix yakale ya rRNA imakhala chidutswa chokhala ndi chingwe chimodzi chotalikirana pafupifupi mzere wowongoka.Kukhalapo kwa zigawo zachilendozi kumapangitsa kuti microsporidia rRNA imange zidutswa zakutali za rRNA pogwiritsa ntchito zigawo zochepa za RNA.
Chitsanzo chochititsa chidwi kwambiri cha kusintha kwachisinthikochi chikhoza kuwonedwa mu H18 25S rRNA helix (mkuyu 3).Mu mitundu yochokera ku E. coli kupita kwa anthu, maziko a rRNA helix ali ndi 24-32 nucleotides, kupanga helix yosadziwika pang'ono.M'mapangidwe a ribosomal omwe amadziwika kale kuchokera ku V. necatrix ndi P. locustae, 31,32 maziko a H18 helix amatsekedwa pang'ono, koma nucleotide base pairing imasungidwa.Komabe, mu E. cuniculi chidutswa cha rRNA ichi chimakhala cholumikizira chachifupi kwambiri 228UUUGU232 ndi 301UUUUUUUU307.Mosiyana ndi zidutswa za rRNA, zolumikizira zokhala ndi uridine sizimazungulira kapena kukhudzana kwambiri ndi mapuloteni a ribosomal.M'malo mwake, amatenga zosungunulira zotseguka komanso zosasunthika momwe zingwe za rRNA zimakulitsidwa pafupifupi mowongoka.Conformation yotambasulayi ikufotokoza momwe E. cuniculi amagwiritsira ntchito maziko a 12 RNA okha kuti akwaniritse kusiyana kwa 33 Å pakati pa H16 ndi H18 rRNA helices, pamene mitundu ina imafuna zosachepera kuwirikiza kawiri rRNA maziko kudzaza kusiyana.
Chifukwa chake, titha kuwonetsa kuti, popinda mopanda mphamvu, ma parasitic microsporidia apanga njira yolumikizira magawo a rRNA omwe amakhalabe otetezedwa ku mitundu yonse ya zamoyo m'magawo atatu amoyo.Mwachiwonekere, posonkhanitsa masinthidwe omwe amasintha ma helice a rRNA kukhala zolumikizira zazifupi za poly-U, E. cuniculi imatha kupanga zidutswa zachilendo za rRNA zomwe zimakhala ndi ma nucleotide ochepa momwe angathere kuti agwirizane ndi tizidutswa ta distal rRNA.Izi zimathandiza kufotokoza momwe microsporidia idakwanitsira kuchepetsedwa kwakukulu kwamapangidwe awo oyambira popanda kutaya kukhulupirika kwawo komanso magwiridwe antchito.
Chinthu china chachilendo cha E. cuniculi rRNA ndi maonekedwe a rRNA popanda thickenings (mkuyu 4).Mabulge ndi ma nucleotide opanda ma awiri awiri oyambira omwe amapotoza kuchokera ku RNA helix m'malo mobisala mmenemo.Ma protrusions ambiri a rRNA amakhala ngati zomatira zamamolekyulu, zomwe zimathandiza kumanga mapuloteni oyandikana ndi ribosomal kapena zidutswa zina za rRNA.Zina mwazotupa zimakhala ngati ma hinges, zomwe zimalola kuti rRNA helix isinthe ndi kupindika bwino kuti apange mapuloteni opangira 41.
a An rRNA protrusion (S. cerevisiae numbering) kulibe mu E. cuniculi ribosome structure, koma ilipo mu eukaryotes ena ambiri b E. coli, S. cerevisiae, H. sapiens, ndi E. cuniculi internal ribosomes.majeremusi alibe zambiri zakale, zotetezedwa kwambiri za rRNA.Kukhuthala kumeneku kumalimbitsa dongosolo la ribosome;Choncho, kusakhalapo kwawo mu microsporidia kumasonyeza kuchepetsedwa kwa bata la rRNA lopinda mu microsporidia tizilombo toyambitsa matenda.Poyerekeza ndi P zimayambira (L7 / L12 zimayambira mu mabakiteriya) zimasonyeza kuti kutaya kwa rRNA tokhala nthawi zina kumagwirizana ndi maonekedwe atsopano pafupi ndi zotayika zotayika.H42 helix mu 23S/28S rRNA ili ndi chotupa chakale (U1206 mu Saccharomyces cerevisiae) chomwe chikuyembekezeka kukhala ndi zaka 3.5 biliyoni zakubadwa chifukwa chotetezedwa m'magawo atatu amoyo.Mu microsporidia, chotupa ichi chimachotsedwa.Komabe, chotupa chatsopano chinawonekera pafupi ndi chotupa chotayika (A1306 mu E. cuniculi).
Chochititsa chidwi, tinapeza kuti E. cuniculi ribosomes alibe zambiri za rRNA zotupa zomwe zimapezeka m'mitundu ina, kuphatikizapo zotupa zoposa 30 zomwe zimasungidwa mu eukaryotes (mkuyu 4a).Kutayika kumeneku kumathetsa kuyanjana kwakukulu pakati pa ma subunits a ribosomal ndi ma helice oyandikana ndi rRNA, nthawi zina kumapanga voids zazikulu zopanda kanthu mkati mwa ribosome, zomwe zimapangitsa E. cuniculi ribosome kukhala porous poyerekeza ndi ribosomes zambiri zachikhalidwe (mkuyu 4b).Mwachidziwitso, tapeza kuti zambiri mwa ziphuphuzi zinatayikanso mu V. necatrix yodziwika kale ndi P. locustae ribosome zomangidwa, zomwe zinanyalanyazidwa ndi kafukufuku wam'mbuyomu31,32.
Nthawi zina kutayika kwa ziphuphu za rRNA kumatsagana ndi kukula kwa ziphuphu zatsopano pafupi ndi zotayika zotayika.Mwachitsanzo, ribosomal P-stem ili ndi chotupa cha U1208 (mu Saccharomyces cerevisiae) chomwe chinapulumuka kuchokera ku E. coli kupita kwa anthu ndipo chifukwa chake chikuyembekezeka kukhala zaka 3.5 biliyoni.Pakupanga mapuloteni, chotupachi chimathandizira tsinde la P kusuntha pakati pa zotseguka ndi zotsekedwa kuti ribosome ipeze zinthu zomasulira ndikuzipereka kumalo omwe akugwira ntchito.Mu E. cuniculi ribosomes, makulidwe awa kulibe;komabe, kuwonjezereka kwatsopano (G883) komwe kumakhala pazigawo zitatu zoyambira kungathandize kuti kubwezeretsanso kusinthika kwabwino kwa P tsinde (Mkuyu 4c).
Deta yathu ya rRNA yopanda ziphuphu imasonyeza kuti kuchepetsa rRNA sikumangokhalira kutayika kwa zinthu za rRNA pamwamba pa ribosome, komanso kungaphatikizepo nucleus ya ribosome, kupanga kachilombo kameneka kamene sikanafotokozedwe m'maselo omasuka.zamoyo zimawonedwa.
Titafanizira mapuloteni ovomerezeka a ribosomal ndi rRNA, tidapeza kuti zida zodziwika bwino za ribosomal sizingathe kufotokozera magawo atatu a chithunzi cha cryo-EM.Awiri mwa zidutswa izi ndi mamolekyu ang'onoang'ono kukula (mkuyu 5, Supplementary mkuyu. 8).Gawo loyamba limayikidwa pakati pa mapuloteni a ribosomal uL15 ndi eL18 pamalo omwe nthawi zambiri amakhala ndi C-terminus ya eL18, yomwe imafupikitsidwa mu E. cuniculi.Ngakhale kuti sitingathe kudziwa kuti molekyuyi ndi yotani, kukula ndi mawonekedwe a chilumba cha kachulukidwechi kumafotokozedwa bwino ndi kukhalapo kwa mamolekyu a spermidine.Kumanga kwake ku ribosome kumakhazikika ndi kusintha kwa microsporidia-specific mu L15 mapuloteni (Asp51 ndi Arg56), omwe amawoneka kuti akuwonjezera kuyanjana kwa ribosome kwa molekyulu yaying'ono iyi, chifukwa amalola uL15 kukulunga kamolekyu kakang'ono kukhala kapangidwe ka ribosomal.Chithunzi chowonjezera 2).8, deta yowonjezera 1, 2).
Kujambula kwa Cryo-EM kusonyeza kukhalapo kwa ma nucleotides kunja kwa ribose ku E. cuniculi ribosome.Mu E. cuniculi ribosome, nucleotide iyi imakhala ndi malo omwewo monga 25S rRNA A3186 nucleotide (Saccharomyces cerevisiae numbering) m'ma ribosomes ena ambiri a eukaryotic.b Mu dongosolo la ribosomal la E. cuniculi, nucleotide iyi ili pakati pa mapuloteni a ribosomal uL9 ndi eL20, motero kukhazikika kukhudzana pakati pa mapuloteni awiriwa.cd eL20 sequence conservation analysis pakati pa microsporidia mitundu.Mtengo wa phylogenetic wa mitundu ya Microsporidia (c) ndi kutsata kangapo kwa mapuloteni a eL20 (d) zimasonyeza kuti zotsalira za nucleotide F170 ndi K172 zimasungidwa mu Microsporidia, kupatula S. lophii, kupatulapo nthambi yoyambirira ya ES39, yomwe imasunga Microsporidia ES39.e Chiwerengerochi chikuwonetsa kuti zotsalira za nucleotide-binding F170 ndi K172 zimapezeka mu eL20 yokha ya microsporidia genome yochepetsedwa kwambiri, koma osati mu eukaryote ina.Zonsezi, deta iyi imasonyeza kuti ma ribosomes a Microsporidian apanga malo omangiriza a nucleotide omwe amawoneka kuti amamanga mamolekyu a AMP ndikuwagwiritsa ntchito kuti akhazikitse mgwirizano wa mapuloteni-mapuloteni mu dongosolo la ribosomal.Kusungidwa kwakukulu kwa malo omangirizawa ku Microsporidia komanso kusapezeka kwake mu eukaryotes ena kukuwonetsa kuti tsambali lingapereke mwayi wosankha kupulumuka kwa Microsporidia.Choncho, thumba la nucleotide-binding mu microsporidia ribosome silikuwoneka kuti ndilowonongeka kapena mawonekedwe a mapeto a kuwonongeka kwa rRNA monga momwe tafotokozera kale, koma ndi njira yothandiza yosinthira yomwe imalola kuti microsporidia ribosome imangirire mwachindunji mamolekyu ang'onoang'ono, kuwagwiritsa ntchito ngati zitsulo zomangira maselo.Zomangamanga za ribosomes.Kupezeka kumeneku kumapangitsa kuti microsporidia ribosome ikhale ribosome yokhayo yomwe imadziwika kuti imagwiritsa ntchito nucleotide imodzi ngati chipika chake chomangira.f Njira yachisinthiko yongopeka yochokera ku kumanga ma nucleotide.
Wachiwiri otsika maselo kulemera kachulukidwe lili pa mawonekedwe pakati ribosomal mapuloteni uL9 ndi eL30 (mkuyu. 5a).Mawonekedwe awa adafotokozedwa kale mu kapangidwe ka Saccharomyces cerevisiae ribosome ngati malo omangira 25S nucleotide ya rRNA A3186 (gawo la ES39L rRNA extension)38.Zinawonetsedwa kuti pakuwonongeka kwa P. locustae ES39L ribosomes, mawonekedwewa amamangiriza nucleotide imodzi yosadziwika 31, ndipo amaganiziridwa kuti nucleotide iyi ndi yochepetsedwa yomaliza ya rRNA, yomwe kutalika kwa rRNA ndi ~ 130-230 maziko.ES39L imachepetsedwa kukhala nucleotide imodzi 32.43.Zithunzi zathu za cryo-EM zimachirikiza lingaliro lakuti kachulukidwe amatha kufotokozedwa ndi ma nucleotides.Komabe, kusamvana kwakukulu kwa kapangidwe kathu kunawonetsa kuti nucleotide iyi ndi molekyulu ya extraribosomal, mwina AMP (Mkuyu 5a, b).
Kenako tinafunsa ngati malo omangira ma nucleotide adawonekera mu E. cuniculi ribosome kapena ngati analipo kale.Popeza kumanga ma nucleotide kumalumikizidwa makamaka ndi zotsalira za Phe170 ndi Lys172 mu eL30 ribosomal protein, tidawunika kusungidwa kwa zotsalirazi mu 4396 oyimira eukaryotes.Monga momwe zilili ndi uL15 pamwambapa, tapeza kuti zotsalira za Phe170 ndi Lys172 zimasungidwa kwambiri mu Microsporidia, koma kulibe mu eukaryotes ena, kuphatikizapo atypical Microsporidia Mitosporidium ndi Amphiamblys, momwe ES39L rRNA fragment siinachepetsedwa (5c 4, 5c4).-e).
Kuphatikizidwa pamodzi, deta iyi ikugwirizana ndi lingaliro lakuti E. cuniculi ndipo mwinamwake ma microsporidia ena ovomerezeka asintha mphamvu yogwira bwino ma metabolites ang'onoang'ono mu dongosolo la ribosome kuti athe kubwezera kuchepa kwa rRNA ndi mapuloteni.Pochita izi, apanga luso lapadera lomanga ma nucleotide kunja kwa ribosome, kusonyeza kuti tizilombo toyambitsa matenda timalipira pogwira ma metabolites ang'onoang'ono ochuluka ndikuwagwiritsa ntchito ngati zotsatsira zowonongeka za RNA ndi zidutswa za mapuloteni..
Gawo lachitatu losayerekezeka la mapu athu a cryo-EM, opezeka mu gawo lalikulu la ribosomal.Kusamvana kwakukulu (2.6 Å) pamapu athu kukuwonetsa kuti kachulukidweko ndi ka mapuloteni okhala ndi zotsalira zazikulu zam'mbali zam'mbali, zomwe zidatipangitsa kuzindikira kachulukidwewa ngati mapuloteni osadziwika kale omwe tidawazindikira kuti Anatchedwa msL2 (Microsporidia- specific protein L2) (njira, chithunzi 6).Kufufuza kwathu kwa homology kunasonyeza kuti msL2 imasungidwa mu Microsporidia clade ya mtundu wa Encephaliter ndi Orosporidium, koma kulibe mu zamoyo zina, kuphatikizapo Microsporidia.Mu dongosolo la ribosomal, msL2 imatenga kusiyana komwe kumapangidwa ndi kutayika kwa ES31L rRNA yowonjezera.Popanda izi, msL2 imathandizira kukhazikika kwa rRNA kupukutira ndipo imatha kubweza kutayika kwa ES31L (Chithunzi 6).
kachulukidwe ka Electron ndi chitsanzo cha Microsporidia-specific ribosomal protein msL2 yopezeka mu E. cuniculi ribosomes.b Ma ribosome ambiri a eukaryotic, kuphatikizapo 80S ribosome of Saccharomyces cerevisiae, ali ndi kukulitsa kwa ES19L rRNA kutayika mu mitundu yambiri ya Microsporidian.Zomwe zidakhazikitsidwa kale za V. necatrix microsporidia ribosome zikusonyeza kuti kutayika kwa ES19L mu tizilombo toyambitsa matenda kulipiridwa ndi kusinthika kwa mapuloteni atsopano a msL1 ribosomal.Mu kafukufukuyu, tapeza kuti E. cuniculi ribosome inapanganso puloteni yowonjezera ya ribosomal RNA mimic monga chipukuta misozi cha kutayika kwa ES19L.Komabe, msL2 (yomwe pano ikufotokozedwa ngati puloteni yongopeka ECU06_1135) ndi msL1 ali ndi zoyambira zosiyanasiyana komanso zachisinthiko.c Kupeza kumeneku kwa m'badwo wa msL1 ndi msL2 ribosomal mapuloteni osagwirizana ndi chisinthiko kukuwonetsa kuti ngati ma ribosomes apeza masinthidwe owononga mu rRNA yawo, amatha kukwanitsa kusiyanasiyana kosiyanasiyana ngakhale kagawo kakang'ono ka mitundu yogwirizana kwambiri.Kupezeka kumeneku kungathandize kumveketsa bwino komwe kunachokera komanso kusinthika kwa ribosome ya mitochondrial, yomwe imadziwika chifukwa cha kuchepa kwake kwa rRNA komanso kusiyanasiyana kwapadziko lonse kwa mapuloteni osiyanasiyana.
Kenaka tinayerekezera mapuloteni a msL2 ndi mapuloteni a msL1 omwe adafotokozedwa kale, mapuloteni okhawo omwe amadziwika kuti ndi microsporidia-specific ribosomal omwe amapezeka mu V. necatrix ribosome.Tinkafuna kuyesa ngati msL1 ndi msL2 ndizogwirizana.Kufufuza kwathu kunasonyeza kuti msL1 ndi msL2 zimakhala ndi msl1 mumtundu wa ribosomal, koma zimakhala ndi zigawo zosiyana za pulayimale ndi zapamwamba, zomwe zimasonyeza chiyambi chawo chodziimira (mkuyu 6).Chifukwa chake, kupezeka kwathu kwa msL2 kumapereka umboni woti magulu amitundu yolumikizana ya eukaryotic amatha kusintha mokhazikika mapuloteni a ribosomal kuti alipire kutayika kwa zidutswa za rRNA.Zomwe anapezazi ndizodziwikiratu kuti ma ribosomes ambiri a cytoplasmic eukaryotic ali ndi mapuloteni osasinthika, kuphatikizapo banja lomwelo la mapuloteni 81 a ribosomal.Maonekedwe a msL1 ndi msL2 m'magulu osiyanasiyana a microsporidia poyankha kutayika kwa magawo otalikirapo a rRNA akuwonetsa kuti kuwonongeka kwa kamangidwe ka maselo a parasite kumapangitsa kuti tizilombo toyambitsa matenda tipeze masinthidwe olipira, zomwe zimatha kupangitsa kuti apezeke m'magulu osiyanasiyana.zomangamanga.
Potsirizira pake, chitsanzo chathu chitatha, tinayerekezera kapangidwe ka E. cuniculi ribosome ndi zimene zinanenedweratu kuchokera ku dongosolo la majeremusi.Mapuloteni angapo a ribosomal, kuphatikizapo eL14, eL38, eL41, ndi eS30, ankaganiziridwa kale kuti akusowa mu E. cuniculi genome chifukwa cha kusakhalapo kwa ma homologue awo ku E. cuniculi genome.Kutayika kwa mapuloteni ambiri a ribosomal kumanenedweratu m'magulu ena ambiri omwe amachepetsedwa kwambiri ndi ma endosymbionts.Mwachitsanzo, ngakhale mabakiteriya ambiri opanda moyo amakhala ndi banja lomwelo la mapuloteni a 54 a ribosomal, 11 okha mwa mabanja omwe ali ndi mapuloteniwa ali ndi ma homologue odziwika mu genome iliyonse yofufuzidwa ya mabakiteriya oletsedwa.Pochirikiza lingaliro ili, kutayika kwa mapuloteni a ribosomal kwawonetsedwa moyesera mu V. necatrix ndi P. locustae microsporidia, omwe alibe mapuloteni a eL38 ndi eL4131,32.
Komabe, mapangidwe athu akuwonetsa kuti eL38, eL41, ndi eS30 okha ndi omwe atayika mu E. cuniculi ribosome.Puloteni ya eL14 idasungidwa ndipo kapangidwe kathu kanawonetsa chifukwa chake puloteni iyi sinapezeke pakufufuza kwa homology (Mkuyu 7).Mu E. cuniculi ribosomes, malo ambiri omangira eL14 amatayika chifukwa cha kuwonongeka kwa rRNA-amplified ES39L.Popanda ES39L, eL14 inataya mawonekedwe ake achiwiri, ndipo 18% yokha ya ndondomeko ya eL14 inali yofanana mu E. cuniculi ndi S. cerevisiae.Kusasungika kotereku ndikodabwitsa chifukwa ngakhale Saccharomyces cerevisiae ndi Homo sapiens - zamoyo zomwe zidasintha zaka 1.5 biliyoni motalikirana - zimagawana zoposa 51% za zotsalira zomwezo mu eL14.Kutayika kodabwitsa kumeneku kumafotokoza chifukwa chake E. cuniculi eL14 pano akufotokozedwa ngati puloteni yoyika M970_061160 osati ngati eL1427 ribosomal protein.
ndi The Microsporidia ribosome inataya ES39L rRNA yowonjezera, yomwe inachotsa pang'ono malo omangira mapuloteni a eL14 ribosomal.Popanda ES39L, puloteni ya eL14 microspore imataya mawonekedwe achiwiri, momwe rRNA-binding α-helix imasintha kukhala loop yochepa.B Mayendedwe angapo akuwonetsa kuti puloteni ya eL14 imasungidwa kwambiri mumitundu ya eukaryotic (57% yotsatizana pakati pa yisiti ndi ma homologues a anthu), koma osasungidwa bwino komanso amasiyana mu microsporidia (momwe zosaposa 24% zotsalira ndizofanana ndi homologue ya eL14).kuchokera ku S. cerevisiae kapena H. sapiens).Kusamalidwa bwino kotsatizanaku komanso kusinthika kwadongosolo lachiwiri kumafotokoza chifukwa chake eL14 homologue sinapezekepo mu E. cuniculi komanso chifukwa chake mapuloteniwa amaganiziridwa kuti adatayika mu E. cuniculi.Mosiyana ndi izi, E. cuniculi eL14 idatchulidwa kale ngati mapuloteni a M970_061160.Kafukufukuyu akusonyeza kuti mitundu yosiyanasiyana ya ma microsporidia genome panopa ndi yochuluka kwambiri: majini ena omwe panopa akuganiziridwa kuti atayika mu microsporidia amasungidwa, ngakhale kuti ali osiyana kwambiri;m'malo mwake, ena amaganiziridwa kuti ali ndi ma gene a microsporidia a mapuloteni enieni a nyongolotsi (mwachitsanzo, mapuloteni ongoyerekeza M970_061160) amayika mapuloteni osiyanasiyana opezeka mu eukaryotes ena.
Kupeza uku kukuwonetsa kuti kusintha kwa rRNA kungayambitse kutayika kwakukulu kwa kasungidwe kakusungidwa m'mapuloteni oyandikana ndi ribosomal, kupangitsa kuti mapuloteniwa asawonekere pakufufuza kwa homology.Motero, tinganene mopambanitsa kuti tinthu tating’ono tating’onoting’ono timene timawonongeka chifukwa cha kuwonongeka kwa maselo, popeza kuti mapuloteni ena amene amaganiziridwa kuti atayika amakhalabe, ngakhale kuti amasinthidwa kwambiri.
Kodi tizilombo tingatani kuti tisungebe kugwira ntchito kwa makina awo a mamolekyu mumkhalidwe wochepa kwambiri wa ma genome?Phunziro lathu limayankha funsoli pofotokoza mmene mamolekyu amachulukira (ribosome) a E. cuniculi, chamoyo chomwe chili ndi tinthu tating’ono kwambiri ta eukaryotic genomes.
Zakhala zikudziwika kwa zaka pafupifupi makumi awiri kuti mapuloteni ndi mamolekyu a RNA mu tizilombo toyambitsa matenda nthawi zambiri amasiyana ndi mamolekyu awo amtundu wamtundu wamtundu wamtundu waufulu chifukwa alibe malo olamulira khalidwe, amachepetsedwa mpaka 50% ya kukula kwawo mu tizilombo toyambitsa matenda, etc.zambiri zofooketsa masinthidwe omwe amalepheretsa kupindika ndi kugwira ntchito.Mwachitsanzo, ma ribosomes a tinthu tating'ono tating'ono tating'onoting'ono, kuphatikiza tizilombo toyambitsa matenda ndi ma endosymbionts, akuyembekezeka kusowa mapuloteni angapo a ribosomal ndi gawo limodzi mwa magawo atatu a rRNA nucleotides poyerekeza ndi zamoyo zaufulu 27, 29, 30, 49.
Kafukufuku wathu akuwonetsa kuti mapangidwe a ma macromolecules amatha kuwulula mbali zambiri za chisinthiko zomwe zimakhala zovuta kuzichotsa kumaphunziro achikhalidwe ofananiza a genomic a tizilombo toyambitsa matenda ndi zamoyo zina zoletsedwa (Zowonjezera mkuyu 7).Mwachitsanzo, chitsanzo cha puloteni ya eL14 chikuwonetsa kuti titha kuwonetsa kuchuluka kwenikweni kwa kuwonongeka kwa zida zama cell mu mitundu ya parasitic.Tizilombo tating'onoting'ono tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono tating'ono ta microsporidia.Komabe, zotsatira zathu zimasonyeza kuti ena mwa majini ooneka ngati enieni amangokhala mitundu yosiyana kwambiri ya majini omwe amapezeka mu eukaryotes ena.Kuphatikiza apo, chitsanzo cha mapuloteni a msL2 chikuwonetsa momwe timanyalanyaza mapuloteni atsopano a ribosomal ndikuchepetsa zomwe zili m'makina a cell a parasitic.Chitsanzo cha mamolekyu ang'onoang'ono chimasonyeza momwe tinganyalanyaze luso lapamwamba kwambiri la maselo a parasitic omwe angawapatse ntchito zatsopano zamoyo.
Kuphatikizidwa pamodzi, zotsatirazi zimathandizira kumvetsetsa kwathu kusiyana pakati pa mamolekyu a zamoyo zomwe zili ndi malire ndi mafananidwe awo mu zamoyo zopanda moyo.Timasonyeza kuti makina a mamolekyu, omwe ankaganiziridwa kale kuti amachepetsedwa, amawonongeka, komanso amatha kusintha masinthidwe osiyanasiyana ofooketsa, m'malo mwake amakhala ndi mawonekedwe achilendo omwe amanyalanyazidwa.
Kumbali ina, zidutswa za rRNA zopanda bulky ndi zidutswa zosakanikirana zomwe tidazipeza mu ribosomes za E. cuniculi zimasonyeza kuti kuchepetsa ma genome kungasinthe ngakhale zigawo za makina oyambirira a maselo omwe amasungidwa m'madera atatu a moyo - pambuyo pa zaka pafupifupi 3.5 biliyoni.chisinthiko chodziyimira pawokha cha zamoyo.
Zidutswa za rRNA zopanda ziphuphu komanso zosakanizika mu E. cuniculi ribosomes ndizosangalatsa kwambiri powunikira maphunziro am'mbuyomu a mamolekyu a RNA mu mabakiteriya a endosymbiotic.Mwachitsanzo, mu aphid endosymbiont Buchnera aphidicola, rRNA ndi mamolekyu a tRNA asonyezedwa kuti ali ndi mapangidwe osagwirizana ndi kutentha chifukwa cha A + T bias komanso kuchuluka kwa magulu awiri omwe si a canonical base20,50.Kusintha kumeneku kwa RNA, komanso kusintha kwa mamolekyu a mapuloteni, tsopano akuganiziridwa kuti ndi chifukwa cha kudalira kwambiri kwa endosymbionts pa okondedwa komanso kulephera kwa endosymbionts kusamutsa kutentha 21, 23.Ngakhale kuti parasitic microsporidia rRNA ili ndi kusintha kosiyana, momwe kusinthaku kumasonyeza kuti kuchepetsa kukhazikika kwa kutentha komanso kudalira kwambiri mapuloteni a chaperone kungakhale zinthu zodziwika bwino za mamolekyu a RNA mu zamoyo zomwe zimakhala ndi majeremusi ochepa.
Kumbali ina, mapangidwe athu amasonyeza kuti tizilombo toyambitsa matenda tizilombo toyambitsa matenda tasintha luso lapadera lolimbana ndi rRNA yosungidwa bwino ndi zidutswa za mapuloteni, kukulitsa luso logwiritsa ntchito ma metabolites ang'onoang'ono ochuluka komanso opezeka mosavuta monga zotsatsira zowonongeka za rRNA ndi zidutswa za mapuloteni.Kuwonongeka kwa mamolekyu..Lingaliro ili limachirikizidwa ndi mfundo yakuti mamolekyu ang'onoang'ono omwe amalipira kutaya kwa zidutswa za mapuloteni mu rRNA ndi ribosomes za E. cuniculi zimamangiriza zotsalira za microsporidia mu uL15 ndi eL30 mapuloteni.Izi zikusonyeza kuti kumanga mamolekyu ang'onoang'ono ku ribosomes kungakhale chinthu chosankhidwa bwino, momwe kusintha kwa Microsporidia-enieni mu mapuloteni a ribosomal asankhidwa kuti athe kuonjezera mgwirizano wa ribosomes kwa mamolekyu ang'onoang'ono, omwe angapangitse kuti tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda.Kupezaku kukuwonetsa luso lanzeru pamapangidwe a ma cell a tizilombo tating'onoting'ono ndipo zimatipatsa kumvetsetsa bwino momwe ma cell a ma parasite amagwirira ntchito ngakhale kusintha kwasintha.
Pakali pano, chizindikiritso cha mamolekyu ang'onoang'onowa sichikudziwika.Sizikudziwika chifukwa chake mawonekedwe a mamolekyu ang'onoang'ono mu ribosomal amasiyana pakati pa mitundu ya microsporidia.Makamaka, sizikudziwika chifukwa chake kumangiriza kwa nucleotide kumawonedwa mu ribosomes E. cuniculi ndi P. locustae, osati mu ribosomes V. necatrix, ngakhale kukhalapo kwa F170 zotsalira mu eL20 ndi K172 mapuloteni a V. necatrix.Kuchotsa uku kungayambitsidwe ndi zotsalira 43 uL6 (zomwe zili moyandikana ndi thumba lomangiriza nucleotide), lomwe ndi tyrosine mu V. necatrix osati threonine mu E. cuniculi ndi P. locustae.Unyolo wonunkhira wam'mbali wa Tyr43 ukhoza kusokoneza kumanga kwa ma nucleotide chifukwa cha kuphatikizika kwa steric.Mwinanso, kuchotsedwa kwa nucleotide yowonekera kungakhale chifukwa cha kutsika kochepa kwa kujambula kwa cryo-EM, komwe kumalepheretsa kutsatiridwa kwa zidutswa za V. necatrix ribosomal.
Kumbali ina, ntchito yathu ikusonyeza kuti kuwola kwa majeremusi kungakhale mphamvu yotulukira.Makamaka, kapangidwe ka E. cuniculi ribosome zikusonyeza kuti imfa ya rRNA ndi zidutswa mapuloteni mu microsporidia ribosome amalenga chisinthiko kuthamanga kuti amalimbikitsa kusintha ribosome dongosolo.Zosiyanasiyanazi zimachitika kutali ndi malo ogwirira ntchito a ribosome ndipo zikuwoneka kuti zimathandizira kukonza (kapena kubwezeretsa) msonkhano wabwino kwambiri wa ribosome womwe ungasokonezedwe ndi kuchepa kwa rRNA.Izi zikuwonetsa kuti kupangika kwakukulu kwa microsporidia ribosome kumawoneka kuti kwasintha kukhala kufunikira kosokoneza ma jini.
Mwina izi zikuwonetsedwa bwino ndi kumanga nucleotide, zomwe sizinawonedwepo mu zamoyo zina mpaka pano.Mfundo yakuti zotsalira za nucleotide zomwe zimamangiriza zimakhalapo mu microsporidia, koma osati mu eukaryotes zina, zimasonyeza kuti malo omwe amamangiriza ma nucleotide sizinthu zotsalira zomwe zikudikirira kutha, kapena malo omaliza a rRNA kuti abwezeretsedwe ku mawonekedwe a nucleotides.M'malo mwake, tsamba ili likuwoneka ngati chinthu chothandiza chomwe chikadatha kusinthika pamasankho angapo abwino.Malo omangira nyukiliya amatha kukhala opangidwa mwachilengedwe: kamodzi ES39L itawonongeka, microsporidia amakakamizika kufunafuna chipukuta misozi kuti abwezeretse mulingo woyenera kwambiri wa ribosome biogenesis pakalibe ES39L.Popeza nyukiliotide iyi imatha kutsanzira ma cell a A3186 nucleotide mu ES39L, molekyulu ya nucleotide imakhala yomangira ribosome, kumangika kwake kumapititsidwa bwino ndi kusintha kwa ma eL30.
Pankhani ya kusintha kwa maselo a tizilombo toyambitsa matenda, kafukufuku wathu akusonyeza kuti mphamvu za Darwin za kusankha kwachilengedwe komanso kusuntha kwa majini a kuwonongeka kwa majeremusi sikumagwira ntchito mofanana, koma kumasinthasintha.Choyamba, kusuntha kwa majini kumachotsa mbali zofunika za biomolecule, kupangitsa kuti chipukuta misozi chikhale chofunikira kwambiri.Pokhapokha majeremusi akakwaniritsa chosowachi kudzera mu kusankha kwachilengedwe kwa Darwin m'pamene ma macromolecules awo adzakhala ndi mwayi wokulitsa mikhalidwe yawo yochititsa chidwi komanso yanzeru.Chofunika kwambiri, kusinthika kwa malo omangira ma nucleotide mu E. cuniculi ribosome kumasonyeza kuti njira yotayika yopezera phindu ya kusintha kwa maselo sikuti imangosintha masinthidwe owononga, koma nthawi zina imapereka ntchito zatsopano pa ma macromolecules a parasitic.
Lingaliro limeneli likugwirizana ndi chiphunzitso cha Sewell Wright cha kusuntha kwa kufanana, chomwe chimanena kuti dongosolo lokhwima la kusankha kwachilengedwe limachepetsa kuthekera kwa zamoyo kupanga zatsopano51,52,53.Komabe, ngati kusuntha kwa majini kumasokoneza masankhidwe achilengedwe, izi zitha kubweretsa kusintha komwe sikungathe kusintha (kapena ngakhale kuwononga) koma kumabweretsa kusintha kwina komwe kumapereka kulimba kwambiri kapena zochitika zatsopano zamoyo.Ndondomeko yathu imachirikiza lingaliroli posonyeza kuti mtundu womwewo wa masinthidwe omwe amachepetsa khola ndi ntchito ya biomolecule akuwoneka kuti ndiye choyambitsa chachikulu cha kusintha kwake.Mogwirizana ndi chitsanzo cha chisinthiko chopambana, kafukufuku wathu amasonyeza kuti kuwonongeka kwa majeremusi, komwe kumawoneka ngati njira yowonongeka, kumakhalanso dalaivala wamkulu wa zatsopano, nthawi zina ndipo mwinamwake ngakhale nthawi zambiri kulola macromolecules kupeza ntchito zatsopano za parasitic.akhoza kuzigwiritsa ntchito.