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ʻO ka hoʻololi ʻana o nā ʻōpala microbial e pili ana i ka counteraction ma waena o ke koho kūlohelohe, ka mea e hoʻomaikaʻi ai i nā parasites, a me ka genetic drift, ka mea e hoʻopau ai nā parasite i nā genes a hōʻiliʻili i nā hoʻololi ʻino.Maʻaneʻi, no ka hoʻomaopopoʻana i keʻano o kēia kū'ēʻana ma ka pālākiō o ka macromolecule hoʻokahi, wehewehe mākou i keʻano cryo-EM o ka ribosome o Encephalitozoon cuniculi, he mea ola eukaryotic me kekahi o nā genome liʻiliʻi loa ma keʻano.ʻO ka hoʻohaʻahaʻa loa o ka rRNA ma E. cuniculi ribosomes ua hele pū ʻia me nā hoʻololi hoʻololi ʻole ʻia, e like me ka hoʻomohala ʻana o nā mea hoʻohui rRNA fused i ʻike ʻole ʻia a me ka rRNA me ka ʻole o nā bulges.Eia kekahi, ua ola ka E. cuniculi ribosome i ka nalowale o nā ʻāpana rRNA a me nā protein ma o ka hoʻomohala ʻana i ka hiki ke hoʻohana i nā moleke liʻiliʻi e like me ke ʻano hoʻohālike o nā ʻāpana rRNA degraded a me nā protein.Ma ke ʻano holoʻokoʻa, hōʻike mākou i nā hale molekala i manaʻo lōʻihi e hoʻemi ʻia, degenerate, a ma lalo o nā hoʻololi hoʻonāwaliwali i loaʻa i nā ʻano hana uku e hoʻomau iā lākou e ʻoi aku ka nui o nā ʻokiʻoki molecular.
Ma muli o ka nui o nā pūʻulu o nā microbial parasites he mau mea hana molekala kūikawā e hoʻohana ai i kā lākou mau pūʻali, pono mākou e hoʻomohala i nā lāʻau lapaʻau like ʻole no nā hui like ʻole o nā parasites1,2.Eia naʻe, hōʻike nā hōʻike hou i kekahi mau ʻano o ka evolution parasite he convergent a ʻike nui ʻia, e hōʻike ana i kahi kumu kūpono no nā interventions therapeutic ākea i nā microbial parasites3,4,5,6,7,8,9.
Ua ʻike ʻia ka hana ma mua i kahi ʻano evolutionary maʻamau i nā microbial parasites i kapa ʻia ka genome reduction a i ʻole genome decay10,11,12,13.Hōʻike ka noiʻi o kēia manawa i ka wā e haʻalele ai nā microorganisms i ko lākou ola manuahi a lilo i intracellular parasites (a i ʻole endosymbionts), hoʻomaka ko lākou genomes i nā metamorphoses lohi akā kupaianaha i nā miliona o nā makahiki9,11.Ma ke kaʻina hana i kapa ʻia ʻo genome decay, hōʻiliʻili nā microbial parasites i nā mutation deleterious e hoʻohuli ai i nā genes koʻikoʻi ma mua i pseudogenes, e alakaʻi ana i ka nalowale ʻana o ka gene a me ka hiolo ʻana o ka mutational14,15.Hiki i kēia hāʻule ke hoʻopau i ka 95% o nā genes i loko o nā mea ola intracellular kahiko loa i hoʻohālikelike ʻia me nā ʻano ola manuahi pili pili.No laila, ʻo ka evolution o intracellular parasites he tug-of-war ma waena o ʻelua mau mana kū'ē: Darwinian koho kūlohelohe, alakaʻi i ka hoʻomaikaʻi ʻana o nā parasites, a me ka hāʻule ʻana o ka genome, e hoʻolei ana i nā parasites i ka poina.Pehea ka puka ʻana mai o ka parasite mai kēia tug-of-war a hoʻopaʻa i ka hana o kona ʻano molekala ʻaʻole maopopo.
ʻOiai ʻaʻole i hoʻomaopopo piha ʻia ke ʻano o ka genome decay, ʻike ʻia ma muli o ka neʻe pinepine ʻana.Ma muli o ka noho ʻana o nā parasites i nā heluna liʻiliʻi, asexual, a me ka genetically, ʻaʻole hiki iā lākou ke hoʻopau maikaʻi i nā hoʻololi deleterious i loaʻa i kekahi manawa i ka wā o ka hana hou ʻana o DNA.Ke alakaʻi nei kēia i ka hōʻiliʻili hiki ʻole o nā mutation ʻino a me ka hōʻemi ʻana o ka genome parasite.ʻO ka hopena, ʻaʻole nalo wale ka parasite i nā genes i pono ʻole no kona ola ʻana i loko o ke kaiapuni intracellular.ʻO ka hiki ʻole o nā heluna parasite ke hoʻopau pono i nā mutations deleterious sporadic e hoʻonui ai i kēia mau hoʻololi i loko o ka genome, me kā lākou mau genes koʻikoʻi.
ʻO ka hapa nui o kā mākou ʻike i kēia manawa e pili ana i ka hoʻemi ʻana o ka genome e hoʻokumu wale ʻia ma ka hoʻohālikelike ʻana i nā kaʻina genome, me ka liʻiliʻi o ka nānā ʻana i nā loli i nā molekole maoli e hana nei i nā hana mālama hale a lilo i mau pahuhopu lāʻau.Ua hōʻike nā haʻawina hoʻohālikelike i ke kaumaha o nā mutation microbial intracellular deleterious e predispose i nā protein a me nā waikawa nucleic i misfold a hōʻuluʻulu, e hoʻonui iā lākou i ka chaperone a me ka hypersensitive i ka wela19,20,21,22,23.Eia kekahi, ʻo nā parasite like ʻole—evolution kūʻokoʻa i hoʻokaʻawale ʻia i kekahi manawa e like me ka 2.5 biliona makahiki-ua ʻike i ka nalowale like ʻana o nā kikowaena mana maikaʻi i kā lākou protein synthesis5,6 a me nā mīkini hoʻoponopono DNA24.Eia nō naʻe, liʻiliʻi ka ʻike e pili ana i ka hopena o ka nohona intracellular i nā waiwai ʻē aʻe a pau o nā macromolecules cellular, me ka hoʻololi ʻana i ka molekala i ka piʻi nui ʻana o nā mutations deleterious.
Ma kēia hana, i mea e hoʻomaopopo maikaʻi ai i ka ulu ʻana o nā protein a me nā waikawa nucleic o nā microorganism intracellular, ua hoʻoholo mākou i ke ʻano o nā ribosomes o ka intracellular parasite Encephalitozoon cuniculi.ʻO ka E. cuniculi he mea ʻano like me ka fungus no ka hui o nā microsporidia parasitic i loaʻa nā genomes eukaryotic liʻiliʻi a no laila ua hoʻohana ʻia ma ke ʻano he kumu hoʻohālike e aʻo ai i ka genome decay25,26,27,28,29,30.I kēia mau lā, ua hoʻoholo ʻia ka hana ribosome cryo-EM no nā genomes i hoʻemi ʻia o Microsporidia, Paranosema locustae, a me Vairimorpha necatrix31,32 (~ 3.2 Mb genome).Hōʻike kēia mau hale e uku ʻia kekahi nalowale o ka hoʻonui ʻana i ka rRNA e ka hoʻomohala ʻana i nā pilina hou ma waena o nā protein ribosomal kokoke a i ʻole ka loaʻa ʻana o nā protein ribosomal msL131,32 hou.Species Encephalitozoon (genome ~ 2.5 miliona bp), a me ko lākou pili pili loa Ordospora, hōʻike i ka degere hope loa o ka genome ho'ēmi i loko o eukaryotes - ua emi iho lakou malalo o 2000 protein-coding genes, a ua manaoia ko lakou ribosomes 'aʻole wale nele i ka rRNA mau'āpana hoʻonui (rRNA mau'āpana e hoʻokaʻawale ai i ko lakou eukaryol ribosomes a me ka nele o ko lakou mau pūmua. homologues i ka E. cuniculi genome26,27,28.No laila, ua hoʻoholo mākou e hiki i ka E. cuniculi ribosome ke hōʻike i nā hoʻolālā i ʻike mua ʻole ʻia no ka hoʻololi ʻana i ka molekala i ka genome decay.
ʻO kā mākou hana cryo-EM e hōʻike ana i ka liʻiliʻi liʻiliʻi eukaryotic cytoplasmic ribosome e hōʻike ʻia a hāʻawi i ka ʻike i ke ʻano o ka hopena hope loa o ka hōʻemi genome i ke ʻano, ka hui ʻana, a me ka hoʻomohala ʻana o ka mīkini molekala i hoʻohui ʻia i ke kelepona.Ua ʻike mākou ua uhaki ka E. cuniculi ribosome i nā loina i mālama nui ʻia o ka RNA folding a me ka hui ribosome, a ua ʻike ʻia kahi pūmua ribosomal hou i ʻike ʻole ʻia.ʻAʻole i manaʻo ʻia, hōʻike mākou ua ulu ka microsporidia ribosomes i ka hiki ke hoʻopaʻa i nā moleke liʻiliʻi, a ke kuhi nei ʻo ka ʻoki ʻana i ka rRNA a me nā protein e hoʻoulu i nā mea hou evolutionary e hāʻawi i nā ʻano pono i ka ribosome.
No ka hoʻomaikaʻi ʻana i ko mākou ʻike i ka ulu ʻana o nā protein a me nā waikawa nucleic i loko o nā mea ola intracellular, ua hoʻoholo mākou e hoʻokaʻawale i nā spores E. cuniculi mai nā moʻomeheu o nā maʻi maʻi maʻi i mea e hoʻomaʻemaʻe ai i kā lākou ribosomes a hoʻoholo i ke ʻano o kēia mau ribosomes.He paʻakikī ka loaʻa ʻana o ka nui o nā microsporidia parasitic no ka mea ʻaʻole hiki ke hoʻoulu ʻia ka microsporidia i kahi ʻano meaʻai.Akā, ulu lākou a hoʻohua wale i loko o ka hale hoʻokipa.No laila, no ka loaʻa ʻana o ka biomass E. cuniculi no ka hoʻomaʻemaʻe ribosome, ua hoʻopōʻino mākou i ka laina cell kidney mammalian RK13 me nā spores E. cuniculi a hoʻomaʻamaʻa i kēia mau maʻi maʻi no kekahi mau pule e hiki ai iā E. cuniculi ke ulu a hoʻonui.Me ka hoʻohana ʻana i kahi monolayer cell ma kahi o ka hapalua mika square, hiki iā mākou ke hoʻomaʻemaʻe ma kahi o 300 mg o Microsporidia spores a hoʻohana iā lākou e hoʻokaʻawale i nā ribosomes.A laila hoʻopau mākou i nā spores i hoʻomaʻemaʻe ʻia me nā pahu aniani a hoʻokaʻawale i nā ribosomes maʻemaʻe me ka hoʻohana ʻana i ka stepwise polyethylene glycol fractionation o nā lysates.Ua ʻae kēia iā mākou e loaʻa ma kahi o 300 µg o E. cuniculi ribosomes maka no ka nānā ʻana.
A laila, hōʻiliʻili mākou i nā kiʻi cryo-EM me ka hoʻohana ʻana i nā laʻana ribosome i loaʻa a hoʻoponopono i kēia mau kiʻi me ka hoʻohana ʻana i nā masks e pili ana i ka subunit ribosomal nui, poʻo subunit liʻiliʻi, a me nā subunit liʻiliʻi.I loko o kēia kaʻina hana, ua hōʻiliʻili mākou i nā kiʻi o kahi 108,000 ribosomal particles a me nā kiʻi cryo-EM i helu me ka hoʻonā o 2.7 Å (Nā Kiʻi Hoʻohui 1-3).A laila, hoʻohana mākou i nā kiʻi cryoEM e hoʻohālike i ka rRNA, ribosomal protein, a me ka hibernation factor Mdf1 pili me E. cuniculi ribosomes (Fig. 1a, b).
He Kapili o ka E. cuniculi ribosome i paʻakikī me ka mea hibernation Mdf1 (pdb id 7QEP).b Map of hibernation factor Mdf1 pili me ka E. cuniculi ribosome.c Ka palapala 'āina hoʻolālā lua e hoʻohālikelike ana i ka rRNA i hoʻihoʻi ʻia ma nā ʻano Microsporidian me nā hale ribosomal i ʻike ʻia.Hōʻike nā paneli i kahi o nā ʻāpana rRNA i hoʻonui ʻia (ES) a me nā wahi hana ribosome, me ka decoding site (DC), ka sarcinicin loop (SRL), a me ka peptidyl transferase center (PTC).d 'O ka mānoanoa electron e pili ana i ke kikowaena peptidyl transferase o ka E. cuniculi ribosome e hō'ike ana he 'ano like ko kēia kahua catalytic i loko o ka E. cuniculi parasite a me kona mau pū'ali, me H. sapiens.e, f ʻO ka mānoanoa electron pili o ke kikowaena decoding (e) a me ka hoʻolālā schematic o ke kikowaena decoding (f) e hōʻike ana he koena ʻo E. cuniculi U1491 ma kahi o A1491 (E. coli helu) i nā eukaryotes he nui.Hōʻike kēia hoʻololi e pili ana paha ʻo E. cuniculi i nā lāʻau antibiotic e kuhikuhi ana i kēia wahi hana.
I ka hoʻokaʻawale ʻana i nā hale i hoʻokumu mua ʻia o V. necatrix a me P. locustae ribosomes (ʻo nā hale ʻelua e hōʻike ana i ka ʻohana microsporidia like ʻo Nosematidae a ua like loa kekahi me kekahi), 31,32 E. cuniculi ribosomes e hana i nā kaʻina hana he nui o ka rRNA a me ka ʻāpana protein.ʻO ka denaturation hou aku (Nā Kiʻi Hoʻohui 4-6).I loko o rRNA, ua komo nā hoʻololi ʻoi loa i ka nalowale piha o ka ʻāpana 25S rRNA i hoʻonui ʻia ES12L a me ka degeneration hapa o nā h39, h41, a me H18 helices (Fig. 1c, Supplementary Fig. 4).Ma waena o nā protein ribosomal, ʻo nā hoʻololi ʻoi loa ka nalowale o ka protein eS30 a me ka pōkole o ka eL8, eL13, eL18, eL22, eL29, eL40, uS3, uS9, uS14, uS17, a me eS7 proteins (Supplementary Figures 4, 5).
No laila, ʻike ʻia ka hōʻemi nui ʻana o nā genomes o Encephalotozoon / Ordospora i loko o kā lākou ribosome: E. cuniculi ribosomes e ʻike i ka nalowale loa o ka ʻike protein i loko o nā ribosomes cytoplasmic eukaryotic e pili ana i ka structural characterization, ʻaʻole lākou i loaʻa i kēlā mau rRNA a me nā ʻāpana protein i mālama nui ʻia i loko o nā eukaryotes ʻekolu wale nō.Hāʻawi ke ʻano o ka E. cuniculi ribosome i ka molecular model mua no kēia mau hoʻololi a hōʻike i nā hanana evolutionary i nānā ʻole ʻia e nā genomics comparative a me nā haʻawina o ka ʻano biomolecular intracellular (Supplementary Fig. 7).Ma lalo nei, wehewehe mākou i kēlā me kēia mau hanana me ko lākou kumu evolutionary a me ko lākou hopena i ka hana ribosome.
A laila ʻike mākou, ma waho aʻe o nā ʻokiʻoki rRNA nui, loaʻa i nā ribosomes E. cuniculi nā ʻano rRNA ma kekahi o kā lākou mau wahi hana.ʻOiai ʻo ka peptidyl transferase kikowaena o ka E. cuniculi ribosome he ʻano like me nā ribosomes eukaryotic ʻē aʻe (Fig. 1d), ʻokoʻa ke kikowaena decoding ma muli o ka hoʻololi o ke kaʻina ma ka nucleotide 1491 (E. coli helu, Fig. 1e, f).He mea koʻikoʻi kēia nānā ʻana no ka mea ʻo ka pae decoding o nā ribosomes eukaryotic maʻamau i loaʻa nā koena G1408 a me A1491 i hoʻohālikelike ʻia me nā koena ʻano bacterial A1408 a me G1491.Hoʻokumu kēia ʻano hoʻololi i ka ʻokoʻa ʻokoʻa o nā ribosomes bacteria a me eukaryotic i ka ʻohana aminoglycoside o nā lāʻau antibiotic ribosomal a me nā molekole liʻiliʻi ʻē aʻe e kuhikuhi ana i ka pūnaewele decoding.Ma ke kahua decoding o ka E. cuniculi ribosome, ua hoʻololi ʻia ke koena A1491 me U1491, hiki ke hana i kahi pilina paʻa kūʻokoʻa no nā molekole liʻiliʻi e huli ana i kēia kahua hana.Loaʻa ka ʻano like ʻole A14901 i nā microsporidia ʻē aʻe e like me P. locustae a me V. necatrix, e hōʻike ana ua pālahalaha ʻia ma waena o nā ʻano microsporidia (Fig. 1f).
Ma muli o ka hoʻokaʻawale ʻia ʻana o kā mākou E. cuniculi ribosome samples mai ka metabolically inactive spores, ua hoʻāʻo mākou i ka palapala cryo-EM o E. cuniculi no ka hoʻopaʻa ʻana i ka ribosome ma lalo o ke kaumaha a i ʻole ka pōloli.Nā kumu hibernation 31,32,36,37, 38. Ua hoʻohālikelike mākou i ke ʻano i hoʻokumu mua ʻia o ka ribosome hibernating me ka palapala cryo-EM o ka E. cuniculi ribosome.No ka hoʻopaʻa ʻana, ua hoʻohana ʻia nā ribosomes S. cerevisiae i ka paʻakikī me ka hibernation factor Stm138, nā ʻūhini ribosomes i paʻakikī me ka Lso232 factor, a me nā ribosomes V. necatrix i paʻakikī me nā mea Mdf1 a me Mdf231.I ka manawa like, ua ʻike mākou i ka nui cryo-EM e pili ana i ke koena kumu Mdf1.E like me ka Mdf1 i hoʻopaʻa ʻia i ka V. necatrix ribosome, ua hoʻopaʻa pū ʻo Mdf1 i ka ribosome E. cuniculi, kahi e poloka ai i ka pūnaewele E o ka ribosome, hiki ke kōkua i ka loaʻa ʻana o nā ribosomes ke lilo nā spores parasite i metabolically inactivation ma ke kino inactivation (Figure 2).).
Hoʻopaʻa ʻo Mdf1 i ka pūnaewele E o ka ribosome, ʻike ʻia e kōkua i ka hoʻopau ʻana i ka ribosome i ka wā e lilo ai nā spores parasite i ka metabolically inactive.I loko o ke ʻano o ka ribosome E. cuniculi, ua ʻike mākou ua hana ʻo Mdf1 i kahi pilina i ʻike ʻole ʻia me ke kumu L1 ribosome, ka ʻāpana o ka ribosome e hoʻomaʻamaʻa i ka hoʻokuʻu ʻana o ka tRNA deacylated mai ka ribosome i ka wā o ka synthesis protein.Hōʻike kēia mau hoʻopili e hoʻokaʻawale ʻo Mdf1 mai ka ribosome me ka hoʻohana ʻana i ka mīkini like me ka deacetylated tRNA, e hāʻawi ana i ka wehewehe ʻana no ka wehe ʻana o ka ribosome iā Mdf1 e hoʻāla hou i ka synthesis protein.
Eia naʻe, ua hōʻike ko mākou hale i kahi pilina ʻike ʻole ma waena o Mdf1 a me ka L1 ribosome wāwae (ka ʻāpana o ka ribosome e kōkua ana i ka hoʻokuʻu ʻana i ka tRNA deacylated mai ka ribosome i ka wā o ka synthesis protein).ʻO ka mea nui, hoʻohana ʻo Mdf1 i nā pilina like me ka ʻāpana kuʻekuʻe o ka mole tRNA deacylated (Fig. 2).Ua hōʻike ʻia kēia ʻano hoʻohālike molekole i ʻike mua ʻole ʻia ua hoʻokaʻawale ʻo Mdf1 mai ka ribosome me ka hoʻohana ʻana i ka mīkini like me ka deacetylated tRNA, e wehewehe ana i ka wehe ʻana o ka ribosome i kēia kumu hibernation e hoʻāla hou i ka synthesis protein.
I ke kūkulu ʻana i ke kŘkohu rRNA, ua ʻike mākou i ka E. cuniculi ribosome i hoʻopili ʻia i nā ʻāpana rRNA, a mākou i kapa ai i ka fused rRNA (Fig. 3).I loko o nā ribosomes e hoʻopaʻa ana i nā kāʻei kapu ʻekolu o ke ola, hoʻopili ʻia ka rRNA i nā hale kahi i hoʻokumu ʻia ai ka hapa nui o nā kumu rRNA a i ʻole nā ​​​​paʻa kumu a paʻa kekahi i kekahi a i ʻole e launa pū me nā protein ribosomal38,39,40.Eia nō naʻe, i loko o E. cuniculi ribosomes, ʻike ʻia nā rRNA i kēia ʻano kuʻuna ma ka hoʻololi ʻana i kekahi o kā lākou mau helices i nā ʻāpana rRNA i wehe ʻia.
ʻO keʻano o ka H18 25S rRNA helix ma S. cerevisiae, V. necatrix, a me E. cuniculi.ʻO ka maʻamau, i loko o nā ribosomes e pili ana i nā ʻāpana ola ʻekolu, ua hoʻopili kēia loulou i kahi RNA helix i loaʻa he 24 a 34 mau koena.I loko o Microsporidia, ʻokoʻa, ua hoʻemi ʻia kēia mea hoʻopili rRNA i ʻelua mau mea hoʻopili waiwai uridine i hoʻokahi wale nō he 12 koena.ʻO ka hapa nui o kēia mau koena e ʻike ʻia i nā solvents.Hōʻike ke kiʻi i ka microsporidia parasitic e hōʻeha i nā loina maʻamau o ka hoʻopili ʻana i ka rRNA, kahi e hui pinepine ʻia ai nā kumu rRNA me nā kumu ʻē aʻe a i ʻole ke komo ʻana i ka pilina rRNA-protein.I loko o ka microsporidia, lawe kekahi mau ʻāpana rRNA i kahi paʻi maikaʻi ʻole, kahi i lilo ai ka rRNA helix mua i ʻāpana kaula hoʻokahi i hoʻolōʻihi ʻia kokoke i kahi laina pololei.ʻO ka hiki ʻana o kēia mau ʻāpana ʻokoʻa e hiki ai i ka microsporidia rRNA ke hoʻopaʻa i nā ʻāpana rRNA mamao me ka hoʻohana ʻana i kahi helu liʻiliʻi o nā kumu RNA.
Hiki ke ʻike ʻia ka hiʻohiʻona ʻoi loa o kēia hoʻololi evolutionary ma ka H18 25S rRNA helix (Fig. 3).I loko o nā ʻano mai E. coli a hiki i ke kanaka, aia nā kumu o kēia rRNA helix he 24-32 nucleotides, e hana ana i kahi helix maʻamau.Ma nā hale ribosomal i ʻike mua ʻia mai V. necatrix a me P. locustae, 31,32 nā kumu o ka H18 helix i wehe ʻole ʻia, akā mālama ʻia ka pilina kumu nucleotide.Eia naʻe, ma E. cuniculi ua lilo kēia ʻāpana rRNA i mea hoʻopili pōkole loa 228UUUGU232 a me 301UUUUUUUUU307.ʻAʻole e like me nā ʻāpana rRNA maʻamau, ʻaʻole e hui pū kēia mau mea hoʻohui i ka uridine a i ʻole e launa pū me nā protein ribosomal.Akā, hoʻohana lākou i ka solvent-open a me ka wehe piha ʻana i nā hale i hoʻolōʻihi ʻia nā kaula rRNA kokoke pololei.Hōʻike kēia hoʻohālikelike kikoʻī pehea e hoʻohana ai ʻo E. cuniculi i nā kumu RNA he 12 wale nō e hoʻopiha i ka ʻāpana 33 Å ma waena o ka H16 a me ka H18 rRNA helices, aʻo nā ʻano ʻē aʻe e koi ma ka liʻiliʻi ʻelua o ka nui o nā kumu rRNA e hoʻopiha i ke āpau.
No laila, hiki iā mākou ke hōʻike, ma o ka hoʻopili ʻana me ka ikaika ʻole, ua hoʻomohala ʻo parasitic microsporidia i kahi hoʻolālā e ʻaelike ai i kēlā mau ʻāpana rRNA i mālama nui ʻia i nā ʻano i loko o nā ʻāpana ʻekolu o ke ola.ʻIke ʻia, ma ka hōʻiliʻili ʻana i nā mutations e hoʻololi i nā helices rRNA i mau loulou poly-U pōkole, hiki iā E. cuniculi ke hana i nā ʻāpana rRNA ʻokoʻa i loaʻa nā nucleotides liʻiliʻi no ka ligation o nā ʻāpana rRNA distal.Kōkua kēia i ka wehewehe ʻana i ka loaʻa ʻana o ka microsporidia i ka hōʻemi nui ʻana i kā lākou ʻano molekala kumu me ka nalowale ʻole o ko lākou kūkulu ʻana a me ka hana pono.
ʻO kekahi hiʻohiʻona maʻamau o E. cuniculi rRNA ke ʻano o ka rRNA me ka ʻole o ka mānoanoa (Fig. 4).ʻO nā bulges nā nucleotides me ka ʻole o nā pae kumu e wili i waho o ka RNA helix ma mua o ka peʻe ʻana i loko.ʻO ka hapa nui o nā protrusions rRNA e hana ma ke ʻano he molecular adhesives, e kōkua ana i ka hoʻopaʻa ʻana i nā protein ribosomal pili a i ʻole nā ​​ʻāpana rRNA ʻē aʻe.ʻO kekahi o nā puʻupuʻu e hana ma ke ʻano he hinges, e ʻae ana i ka rRNA helix e lohi a pelu maikaʻi loa no ka synthesis protein huahua 41.
a ʻAʻole ʻike ʻia kahi ʻano rRNA (S. cerevisiae numbering) mai ka E. cuniculi ribosome structure, akā aia i loko o ka hapa nui o nā eukaryotes b E. coli, S. cerevisiae, H. sapiens, a me E. cuniculi internal ribosomes.ʻAʻole i loaʻa i nā parasites ka nui o nā puʻupuʻu rRNA kahiko i mālama nui ʻia.Hoʻopaʻa kēia mau mānoanoa i ke ʻano ribosome;no laila, ʻo ko lākou nele i ka microsporidia e hōʻike ana i ka emi ʻana o ka paʻa ʻana o ka hoʻopili ʻana o rRNA i nā parasite microsporidia.ʻO ka hoʻohālikelike ʻana me nā kumu P (L7/L12 stems in bacteria) hōʻike i ka nalowale ʻana o nā puʻupuʻu rRNA i kekahi manawa me ke ʻano o nā puʻupuʻu hou e pili ana i nā puʻupuʻu nalowale.He bulge kahiko ka H42 helix ma ka 23S/28S rRNA (U1206 in Saccharomyces cerevisiae) ma kahi o 3.5 biliona makahiki ma muli o kona pale ʻana i ʻekolu mau wahi o ke ola.I ka microsporidia, hoʻopau ʻia kēia puʻupuʻu.Eia naʻe, ua ʻike ʻia kahi puʻupuʻu hou ma ka ʻaoʻao o ka puʻu nalowale (A1306 ma E. cuniculi).
ʻO ka mea kupanaha, ua ʻike mākou ʻaʻole i nele ka nui o nā puʻupuʻu rRNA i loko o nā ʻano ʻano ʻē aʻe, me ka nui o 30 mau puʻupuʻu i mālama ʻia i loko o nā eukaryotes ʻē aʻe (Fig. 4a).Hoʻopau kēia poho i ka nui o nā pilina ma waena o nā ʻāpana ribosom a me nā helices rRNA e pili ana, i kekahi manawa e hana ana i nā puka ākea nui i loko o ka ribosome, e ʻoi aku ka porous o ka E. cuniculi ribosome ke hoʻohālikelike ʻia me nā ribosomes kahiko (Fig. 4b).No ka mea, ua ʻike mākou ua nalowale ka hapa nui o kēia mau puʻupuʻu i ka V. necatrix a me P. locustae ribosome kūkulu ʻia, i nānā ʻole ʻia e nā ananaly structural mua31,32.
I kekahi manawa ua hele pū ka nalo ʻana o nā puʻupuʻu rRNA me ka ulu ʻana o nā puʻupuʻu hou e pili ana i ka puʻupuʻu nalowale.No ka laʻana, aia i loko o ka ribosomal P-stem kahi puʻupuʻu U1208 (ma Saccharomyces cerevisiae) i ola mai E. coli a i ke kanaka a no laila ua manaʻo ʻia he 3.5 biliona makahiki.I ka wā o ka hana pūmua, kōkua kēia puʻupuʻu i ka neʻe ʻana o ke kumu P ma waena o nā conformations hāmama a pani ʻia i hiki i ka ribosome ke kiʻi i nā mea unuhi a hoʻouna iā lākou i ke kahua paʻa.I loko o E. cuniculi ribosomes, ʻaʻole kēia mānoanoa;akā naʻe, hiki i kahi mānoanoa hou (G883) i loaʻa i loko o ʻekolu mau papa kumu ke kōkua i ka hoʻihoʻi ʻana i ka maʻalahi o ke kumu P (Fig. 4c).
Hōʻike kā mākou ʻikepili i ka rRNA me ka ʻole o ka puʻupuʻu ʻaʻole i kaupalena ʻia ka hōʻemi ʻana o ka rRNA i ka nalowale o nā mea rRNA ma ka ʻili o ka ribosome, akā hiki ke komo pū i ka ribosome nucleus, e hana ana i kahi kīnā molecular parasite-specific i wehewehe ʻole ʻia i loko o nā pūnaewele ola manuahi.ʻike ʻia nā ʻano mea ola.
Ma hope o ka hoʻohālike ʻana i nā protein ribosomal canonical a me ka rRNA, ua ʻike mākou ʻaʻole hiki i nā ʻāpana ribosomal maʻamau ke wehewehe i nā ʻāpana ʻekolu o ke kiʻi cryo-EM.ʻElua o kēia mau ʻāpana ʻāpana liʻiliʻi i ka nui (Fig. 5, Supplementary Fig. 8).Hoʻopili ʻia ka ʻāpana mua ma waena o nā protein ribosomal uL15 a me eL18 i kahi kūlana maʻamau e noho ʻia e ka C-terminus o eL18, i pōkole ʻia ma E. cuniculi.ʻOiai ʻaʻole hiki iā mākou ke hoʻoholo i ke ʻano o kēia mole, ua wehewehe maikaʻi ʻia ka nui a me ke ʻano o kēia mokupuni paʻa i ka loaʻa ʻana o nā molekole spermidine.Hoʻopaʻa ʻia kona hoʻopaʻa ʻana i ka ribosome e nā microsporidia-specific mutations i loko o nā protein uL15 (Asp51 a me Arg56), me he mea lā e hoʻonui i ka pili o ka ribosome no kēia mole liʻiliʻi, no ka mea e ʻae lākou i ka uL15 e hoʻopili i ka mole liʻiliʻi i loko o kahi ʻano ribosomal.Helu 2).8, ikepili hou 1, 2).
ʻO ke kiʻi Cryo-EM e hōʻike ana i ka hele ʻana o nā nucleotides ma waho o ka ribose i hoʻopaʻa ʻia i ka ribosome E. cuniculi.I loko o ka E. cuniculi ribosome, noho kēia nucleotide i kahi like me ka 25S rRNA A3186 nucleotide (Saccharomyces cerevisiae helu) ma ka hapa nui o nā ribosomes eukaryotic.b I loko o ka ribosomal structure o E. cuniculi, aia kēia nucleotide ma waena o nā protein ribosomal uL9 a me eL20, a laila hoʻopaʻa i ka pilina ma waena o nā protein ʻelua.cd eL20 kaʻina mālama mālama i waena o nā ʻano microsporidia.ʻO ka lāʻau phylogenetic o Microsporidia species (c) a me ka hoʻonohonoho ʻana i nā kaʻina he nui o ka protein eL20 (d) e hōʻike ana i nā koena paʻa nucleotide F170 a me K172 i mālama ʻia i loko o ka Microsporidia maʻamau, koe naʻe ʻo S. lophii, koe ka Microsporidia lālā mua, ka mea i mālama i ka ES39L rRNA.e Hōʻike kēia kiʻi i nā koena paʻa nucleotide F170 a me K172 aia wale nō ma eL20 o ka genome microsporidia i hoʻemi nui ʻia, akā ʻaʻole i loko o nā eukaryotes ʻē aʻe.Ma ke ʻano holoʻokoʻa, hōʻike kēia mau ʻikepili ua kūkulu ʻo Microsporidian ribosomes i kahi kahua hoʻopaʻa nucleotide e ʻike ʻia e hoʻopaʻa i nā molekala AMP a hoʻohana iā lākou e hoʻopaʻa i ka pilina protein-protein i loko o ka ribosomal structure.ʻO ka mālama kiʻekiʻe o kēia pūnaewele paʻa i Microsporidia a me kona nele i nā eukaryotes ʻē aʻe e hōʻike ana e hāʻawi paha kēia pūnaewele i kahi pono ola koho no Microsporidia.No laila, ʻaʻole ʻike ʻia ka ʻeke nucleotide-binding i loko o ka microsporidia ribosome he hiʻohiʻona degenerate a i ʻole ke ʻano hopena o ka hoʻohaʻahaʻa ʻana o rRNA e like me ka mea i wehewehe mua ʻia, akā he mea hou evolutionary pono e hiki ai i ka microsporidia ribosome ke hoʻopaʻa pololei i nā molekole liʻiliʻi, me ka hoʻohana ʻana iā lākou e like me nā poloka kūkulu hale.hale kūkulu no nā ribosomes.ʻO kēia ʻike ʻana i ka microsporidia ribosome ka ribosome wale nō i ʻike ʻia e hoʻohana i hoʻokahi nucleotide ma ke ʻano he hale kūkulu hale.f Ke ala hoʻomohala hypothetical i loaʻa mai ka paʻa nucleotide.
Aia ka lua haʻahaʻa haʻahaʻa haʻahaʻa molekala kaumaha ma ka pilina ma waena o ribosomal proteins uL9 a me eL30 (Fig. 5a).Ua wehewehe mua ʻia kēia interface ma ke ʻano o ka Saccharomyces cerevisiae ribosome ma ke ʻano he kahua paʻa no ka nucleotide 25S o rRNA A3186 (kahi o ka hoʻonui ES39L rRNA)38.Ua hōʻike ʻia i loko o nā ribosomes P. locustae ES39L degenerate, ua hoʻopaʻa ʻia kēia interface i kahi nucleotide 31 i ʻike ʻole ʻia, a ua manaʻo ʻia ʻo kēia nucleotide kahi ʻano hope loa o rRNA, kahi o ka lōʻihi o rRNA he ~130-230 kumu.Hoʻemi ʻia ka ES39L i hoʻokahi nucleotide 32.43.Kākoʻo kā mākou mau kiʻi cryo-EM i ka manaʻo e hiki ke wehewehe ʻia ka density e nā nucleotides.Eia naʻe, ʻo ka hoʻonā kiʻekiʻe o kā mākou hale i hōʻike i kēia nucleotide he moleke extraribosomal, ʻo AMP paha (Fig. 5a, b).
A laila, nīnau mākou inā ua ʻike ʻia ka pae hoʻopaʻa nucleotide i ka E. cuniculi ribosome a i ʻole ma mua.No ka mea ua hoʻopili nui ʻia ka paʻa nucleotide e nā koena Phe170 a me Lys172 i loko o ka protein ribosomal eL30, ua loiloi mākou i ka mālama ʻana i kēia mau koena ma 4396 eukaryotes representative.E like me ka hihia o uL15 ma luna, ua ʻike mākou ua mālama nui ʻia nā koena Phe170 a me Lys172 wale nō i ka Microsporidia maʻamau, akā ʻaʻole i loko o nā eukaryotes ʻē aʻe, me atypical Microsporidia Mitosporidium a me Amphiamblys, kahi i hoʻemi ʻole ʻia ka ʻāpana ES39L rRNA 44, 45, 45c.-e).
Hoʻohui pū ʻia, kākoʻo kēia mau ʻikepili i ka manaʻo ua hoʻomohala ʻo E. cuniculi a me nā microsporidia canonical ʻē aʻe i ka hiki ke hopu pono i nā helu he nui o nā metabolites liʻiliʻi i loko o ke ʻano ribosome e uku ai i ka emi ʻana o ka rRNA a me nā pae protein.I ka hana ʻana pēlā, ua hoʻomohala lākou i kahi mana kūʻokoʻa e hoʻopaʻa i nā nucleotides ma waho o ka ribosome, e hōʻike ana i ka hoʻopiʻi ʻana o nā molekala parasitic i ka hopu ʻana i nā metabolites liʻiliʻi nui a me ka hoʻohana ʻana iā lākou e like me ke ʻano o ka RNA degraded a me nā ʻāpana protein..
ʻO ke kolu o ka ʻāpana unsimulated o kā mākou palapala cryo-EM, i loaʻa i ka subunit ribosomal nui.ʻO ka hoʻonā kiʻekiʻe (2.6 Å) o kā mākou palapala 'āina e hōʻike ana no nā protein me nā hui like ʻole o nā koena kaulahao ʻaoʻao nui, i ʻae iā mākou e ʻike i kēia ʻano like me kahi protein ribosomal i ʻike mua ʻia a mākou i ʻike ai Ua kapa ʻia ʻo ia ʻo msL2 (Microsporidia- specific protein L2) (nā ala, helu 6).Ua hōʻike ʻia kā mākou ʻimi homology ua mālama ʻia ka msL2 ma ka ʻāpana Microsporidia o ka genus Encephaliter a me Orosporidium, akā ʻaʻole i loaʻa i nā ʻano ʻano ʻē aʻe, me nā Microsporidia ʻē aʻe.Ma ka ribosomal structure, noho ka msL2 i kahi āpau i hana ʻia e ka nalowale o ka ES31L rRNA lōʻihi.Ma kēia hakahaka, kōkua ʻo msL2 i ka hoʻopaʻa ʻana i ka pelu ʻana i ka rRNA a hiki ke hoʻopaʻi i ka nalowale o ES31L (Figure 6).
he Electron density a me ke kŘkohu o ka Microsporidia-specific ribosomal protein msL2 i loaʻa i loko o E. cuniculi ribosomes.b ʻO ka hapa nui o nā ribosomes eukaryotic, me ka ribosome 80S o Saccharomyces cerevisiae, ua nalowale ka hoʻonui ʻana o ES19L rRNA i ka hapa nui o nā ʻano Microsporidian.ʻO ka mea i hoʻokumu muaʻia o ka V. necatrix microsporidia ribosome e hōʻike ana i ka liloʻana o ES19L i loko o kēia mau parasites i ukuʻia e ka evolution o ka msL1 ribosomal protein hou.I loko o kēia haʻawina, ua ʻike mākou ua hoʻomohala ʻo E. cuniculi ribosome i kahi ribosomal RNA mimic protein ma ke ʻano he uku ʻike no ka nalowale o ES19L.Eia nō naʻe, ʻo msL2 (i hōʻike ʻia i kēia manawa ma ke ʻano he hypothetical ECU06_1135 protein) a me msL1 he ʻokoʻa ka hoʻokumu ʻana a me nā kumu hoʻomohala.ʻO kēia ʻike ʻana o ka hanauna o nā pūmua ribosomal evolutionarily pili ʻole msL1 a me msL2 e hōʻike ana inā hōʻiliʻili nā ribosomes i nā hoʻololi ʻino i kā lākou rRNA, hiki iā lākou ke hoʻokō i nā pae like ʻole o ka ʻokoʻa haku mele ma kahi ʻāpana liʻiliʻi o nā ʻano pili pili.Hiki i kēia ʻike ke kōkua i ka wehewehe ʻana i ke kumu a me ka ulu ʻana o ka ribosome mitochondrial, ka mea i ʻike ʻia no kāna rRNA i hoʻemi nui ʻia a me nā ʻano like ʻole o ka hoʻohui ʻana i ka protein ma waena o nā ʻano.
A laila hoʻohālikelike mākou i ka protein msL2 me ka protein msL1 i wehewehe mua ʻia, ʻo ia wale nō ka microsporidia-specific ribosomal protein i loaʻa i ka V. necatrix ribosome.Makemake mākou e hoʻāʻo inā pili ʻia ka msL1 a me msL2.Ua hōʻike kā mākou hōʻike i ka msL1 a me ka msL2 e noho ana i ka lua like i loko o ka ribosomal structure, akā loaʻa nā ʻano kumu mua a me ke kula kiʻekiʻe, e hōʻike ana i ko lākou kumu kūʻokoʻa evolutionary (Fig. 6).No laila, ʻo kā mākou ʻike ʻana i ka msL2 e hāʻawi i nā hōʻike e hiki i nā pūʻulu o nā ʻano eukaryotic compact ke hoʻololi kūʻokoʻa i nā protein ribosomal kūʻokoʻa e uku ai i ka nalowale o nā ʻāpana rRNA.ʻIke ʻia kēia ʻike ʻana i ka hapa nui o nā ribosomes eukaryotic cytoplasmic i loko o ka protein invariant, me ka ʻohana like o 81 ribosomal proteins.ʻO ka hiʻohiʻona o msL1 a me msL2 ma nā ʻāpana like ʻole o ka microsporidia i ka pane ʻana i ka nalo ʻana o nā ʻāpana rRNA lōʻihi e hōʻike ana i ka hoʻohaʻahaʻa ʻana o ka molecular architecture o ka parasite ke kumu e ʻimi ai nā parasite i nā hoʻololi hoʻololi, a hiki ke alakaʻi i kā lākou loaʻa ʻana i nā heluna parasite like ʻole.nā hale.
ʻO ka hope, i ka pau ʻana o kā mākou kumu hoʻohālike, ua hoʻohālikelike mākou i ke ʻano o ka E. cuniculi ribosome me ka mea i wānana ʻia mai ke kaʻina genome.Ua manaʻo mua ʻia ua nalowale kekahi mau protein ribosomal, me eL14, eL38, eL41, a me eS30 mai ka E. cuniculi genome ma muli o ka ʻike ʻole ʻia o kā lākou homologues mai ka genome E. cuniculi.Ua wānana ʻia ka nalowale ʻana o ka nui o nā protein ribosomal i ka hapa nui o nā parasite intracellular i hoʻemi nui ʻia a me nā endosymbionts.No ka laʻana, ʻoiai ʻo ka hapa nui o nā huakini ola manuahi ka ʻohana hoʻokahi o 54 ribosomal proteins, he 11 wale nō o kēia mau ʻohana protein i ʻike ʻia i nā homologues i ʻike ʻia i kēlā me kēia genome i hoʻopaʻa ʻia.I ke kākoʻo ʻana i kēia manaʻo, ua ʻike ʻia ka nalowale o nā protein ribosomal ma V. necatrix a me P. locustae microsporidia, i nele i nā protein eL38 a me eL4131,32.
Eia naʻe, hōʻike kā mākou mau hale i ka eL38, eL41, a me eS30 wale nō i nalowale i loko o ka E. cuniculi ribosome.Ua mālama ʻia ka protein eL14 a ua hōʻike ko mākou hale i ke kumu i hiki ʻole ai ke loaʻa kēia protein i ka huli homology (Fig. 7).I loko o E. cuniculi ribosomes, ua nalowale ka hapa nui o ka pūnaewele ho'opa'a eL14 ma muli o ka ho'oha'aha'a 'ia o ka rRNA-amplified ES39L.I ka loaʻa ʻole o ES39L, ua nalowale ka eL14 i ka hapa nui o kāna ʻano lua, a ʻo 18% wale nō o ke kaʻina eL14 i like me E. cuniculi a me S. cerevisiae.He mea kupanaha ka mālama ʻana i kēia kaʻina hana maikaʻi ʻole no ka mea ʻo Saccharomyces cerevisiae a me Homo sapiens—nā mea ola i ulu ma 1.5 biliona makahiki kaʻawale—ua ʻoi aku ma mua o 51% o nā koena like ma eL14.Hōʻike ʻia kēia nalowale ʻole o ka mālama ʻana i ke kumu i helu ʻia ai ʻo E. cuniculi eL14 i kēia manawa ma ke ʻano he pūmua M970_061160 putative a ʻaʻole like me ka protein ribosomal eL1427.
a Ua nalowale ka Microsporidia ribosome i ka hoʻonui ʻia ʻana o ES39L rRNA, kahi i hoʻopau ʻokoʻa i ka pūnaewele eL14 ribosomal protein binder.I ka loaʻa ʻole o ES39L, ua nalowale ka eL14 microspore protein i ke ʻano lua, kahi e hoʻohaʻahaʻa ai ka rRNA-binding α-helix i loko o kahi loop loop liʻiliʻi.Hōʻike ʻia ka hoʻohālikelike ʻana i nā kaʻina he nui i mālama nui ʻia ka protein eL14 i nā ʻano eukaryotic (57% sequence identity ma waena o ka hū a me nā homologues kanaka), akā mālama maikaʻi ʻole a ʻokoʻa i ka microsporidia (ʻaʻole i ʻoi aku ma mua o 24% o nā koena i like me ka eL14 homologue).mai S. cerevisiae a i ʻole H. sapiens).ʻO kēia ʻano mālama ʻana i ke kaʻina hana maikaʻi ʻole a me ke ʻano like ʻole o ka hoʻonohonoho lua e wehewehe i ke kumu i ʻike ʻole ʻia ai ka homologue eL14 ma E. cuniculi a no ke aha i manaʻo ʻia ua nalowale kēia pūmua ma E. cuniculi.I ka hoʻokaʻawale ʻana, ua hōʻike mua ʻia ʻo E. cuniculi eL14 ma ke ʻano he protein M970_061160 putative.Ke hōʻike nei kēia ʻike ʻana i ka manaʻo nui o ka microsporidia genome diversity i kēia manawa: mālama ʻia kekahi mau genes i manaʻo ʻia e nalowale i ka microsporidia i kēia manawa, ʻoiai ma nā ʻano ʻokoʻa loa;akā, manaʻo ʻia kekahi e hoʻopaʻa inoa no nā genes microsporidia no nā protein ilo-specific (e laʻa, ka protein hypothetical M970_061160) e hoʻopaʻa inoa maoli no nā protein ʻokoʻa loa i loaʻa i nā eukaryotes ʻē aʻe.
Hōʻike kēia ʻike e hiki i ka denaturation rRNA ke alakaʻi i ka nalowale nui o ka mālama ʻana i ke kaʻina i nā protein ribosomal pili, e hoʻolilo i kēia mau protein i ʻike ʻole ʻia no ka ʻimi homology.No laila, hiki iā mākou ke hoʻomāhuahua i ke kiʻekiʻe maoli o ka hoʻohaʻahaʻa ʻana i ka molekala i loko o nā meaola genome liʻiliʻi, no ka mea, ke mau nei kekahi mau protein i manaʻo ʻia e nalowale, ʻoiai ma nā ʻano i hoʻololi ʻia.
Pehea e hiki ai i nā parasites ke hoʻomau i ka hana o kā lākou mīkini molekala ma lalo o nā kūlana o ka hōʻemi genome loa?Pane kā mākou haʻawina i kēia nīnau ma ka wehewehe ʻana i ke ʻano molekula paʻakikī (ribosome) o E. cuniculi, he mea ola me kekahi o nā genomes eukaryotic liʻiliʻi loa.
Uaʻikeʻia no ka aneaneʻelua mau makahiki heʻano likeʻole ka protein a me ka RNA i loko o ka microbial parasites mai kā lākou mau mole homologous i nāʻano ola manuahi no ka meaʻaʻole lākou i nā kikowaena mana maikaʻi, ua hoʻemiʻia i ka 50% o ko lākou nui i nā microbes olaʻole, etc.nui nā hoʻololi hoʻonāwaliwali e hoʻopilikia i ka pelu ʻana a me ka hana.No ka laʻana, ʻo nā ribosomes o nā meaola genome liʻiliʻi, me ka nui o nā parasites intracellular a me nā endosymbionts, ua manaʻo ʻia e nele i kekahi mau protein ribosomal a hiki i ka hapakolu o nā nucleotides rRNA i hoʻohālikelike ʻia me nā ʻano ola manuahi 27, 29, 30, 49.
Hōʻike kā mākou haʻawina e hiki i ke ʻano o nā macromolecules ke hōʻike i nā ʻano he nui o ka evolution i paʻakikī ke unuhi ʻia mai nā haʻawina genomic comparative kuʻuna o nā parasite intracellular a me nā mea ola ʻē aʻe i hoʻopaʻa ʻia (Supplementary Fig. 7).No ka laʻana, hōʻike ka laʻana o ka protein eL14 hiki iā mākou ke hoʻomāhuahua i ke kiʻekiʻe maoli o ka hoʻohaʻahaʻa ʻana o ka mea molekala i nā ʻano parasitic.Ua manaʻoʻiʻo ʻia he mau haneli o nā ʻano microsporidia-specific genes i nā parasite encephalitic.Eia nō naʻe, hōʻike kā mākou hopena i kekahi o kēia mau genes i ʻike ʻia he mau ʻano ʻokoʻa like ʻole o nā genes i maʻamau i nā eukaryotes ʻē aʻe.Eia kekahi, hōʻike ka laʻana o ka protein msL2 i ko mākou nānā ʻana i nā protein ribosomal hou a hoʻohaʻahaʻa i ka ʻike o nā mīkini molekala parasitic.Hōʻike ka laʻana o nā molekala liʻiliʻi pehea e hiki ai iā mākou ke nānā aku i nā mea hou o ka naʻauao i nā ʻano molekala parasitic e hiki ke hāʻawi iā lākou i ka hana olaola hou.
Hoʻohui pū ʻia, hoʻomaikaʻi kēia mau hopena i ko mākou ʻike ʻana i nā ʻokoʻa ma waena o nā ʻano molekala o nā mea i hoʻopaʻa ʻia i ka hoʻokipa a me ko lākou mau hoa i nā mea ola manuahi.Hōʻike mākou i nā mīkini molekala lōʻihi i manaʻo ʻia e hoʻemi ʻia, degenerate, a ma lalo o nā ʻano hoʻololi hoʻonāwaliwali like ʻole, ma kahi o nā hiʻohiʻona ʻokoʻa i nānā ʻole ʻia.
Ma kekahi ʻaoʻao, ʻo nā ʻāpana rRNA non-bulky a me nā ʻāpana fused i loaʻa iā mākou i loko o nā ribosomes o E. cuniculi e hōʻike ana e hiki ke hoʻololi i ka genome hōʻemi hiki ke hoʻololi i kēlā mau ʻāpana o ka mīkini molekele kumu i mālama ʻia i loko o nā ʻāpana ʻekolu o ke ola - ma hope o 3.5 biliona makahiki.kūʻokoʻa evolution o nā ʻano.
ʻO nā ʻāpana rRNA bulge-free a fused i loko o E. cuniculi ribosomes he mea hoihoi nui i ka mālamalama o nā haʻawina mua o nā molekala RNA i loko o ka bacteria endosymbiotic.No ka laʻana, i loko o ka aphid endosymbiont Buchnera aphidicola, rRNA a me tRNA molekele ua hōʻike 'ia i ka wela-koʻikoʻi hale ma muli o ka A + T haku mele 'ana a me ka kiʻekiʻe o ka non-canonical kumu pairs20,50.ʻO kēia mau hoʻololi i ka RNA, a me ka hoʻololi ʻana i nā molekole protein, i kēia manawa ke manaʻo ʻia he kuleana no ka overdependence o endosymbionts i nā hoa a me ka hiki ʻole o nā endosymbionts ke hoʻololi i ka wela 21, 23.ʻOiai he mau loli like ʻole ka microsporidia rRNA, ʻo ke ʻano o kēia mau hoʻololi e hōʻike ana i ka emi ʻana o ke kūpaʻa wela a me ka hilinaʻi kiʻekiʻe ma luna o nā protein chaperone he mau hiʻohiʻona maʻamau o nā molekala RNA i loko o nā meaola me nā genome hoʻemi.
Ma ka ʻaoʻao ʻē aʻe, hōʻike kā mākou mau hale i ka microsporidia parasite i hoʻomohala i kahi mana kūʻokoʻa e kūʻē i ka rRNA i mālama nui ʻia a me nā ʻāpana protein, e hoʻomohala ana i ka hiki ke hoʻohana i nā metabolites liʻiliʻi a maʻalahi e like me ke ʻano hoʻohālike o ka rRNA degenerate a me nā ʻāpana protein.ʻO ka hoʻohaʻahaʻa ʻana i ka molecular structure..Kākoʻo ʻia kēia manaʻo e ka ʻoiaʻiʻo o nā molekala liʻiliʻi e uku ai i ka nalowale o nā ʻāpana protein i loko o ka rRNA a me nā ribosomes o E. cuniculi e hoʻopaʻa i nā koena microsporidia-specific i loko o nā protein uL15 a me eL30.Hōʻike kēia i ka hoʻopaʻa ʻana o nā molekole liʻiliʻi i nā ribosomes he huahana ia o ka koho maikaʻi, kahi i koho ʻia ai nā hoʻololi ʻokoʻa Microsporidia i nā protein ribosomal no ko lākou hiki ke hoʻonui i ka pili o nā ribosomes no nā molekole liʻiliʻi, hiki ke alakaʻi i nā mea ola ribosomal ʻoi aku ka maikaʻi.Hōʻike ka ʻike i kahi mea hou akamai i loko o ke ʻano molecular o nā microbial parasites a hāʻawi iā mākou i ka ʻike maikaʻi aʻe i ke ʻano o ka mālama ʻana o nā molecular parasite i kā lākou hana ʻoiai ʻo ka reductive evolution.
I kēia manawa, ʻaʻole maopopo ka ʻike ʻana o kēia mau molekala liʻiliʻi.ʻAʻole maopopo i ke kumu i ʻokoʻa ai ke ʻano o kēia mau moleke liʻiliʻi i loko o ke ʻano ribosomal ma waena o nā ʻano microsporidia.ʻO keʻano,ʻaʻole maopopo i ke kumu iʻikeʻia ai ka paʻa nucleotide i loko o nā ribosomes o E. cuniculi a me P. locustae, aʻaʻole i loko o nā ribosomes o V. necatrix, me ka loaʻaʻana o ke koena F170 i nā protein eL20 a me K172 o V. necatrix.Hiki ke hoʻopau ʻia kēia holoi ʻana ma muli o ke koena 43 uL6 (aia e pili ana i ka ʻeke hoʻopaʻa nucleotide), ʻo ia ka tyrosine ma V. necatrix a ʻaʻole threonine ma E. cuniculi a me P. locustae.Hiki i ke kaulahao ʻaoʻao ʻala nui o Tyr43 ke hoʻopilikia i ka hoʻopaʻa ʻana o ka nucleotide ma muli o ka overlap steric.ʻO kahi ʻē aʻe, ʻo ka holoi ʻana o ka nucleotide ma muli o ka haʻahaʻa haʻahaʻa o ke kiʻi cryo-EM, kahi e keʻakeʻa ai i ka hoʻohālike ʻana o nā ʻāpana ribosomal V. necatrix.
Ma ka ʻaoʻao ʻē aʻe, hōʻike kā mākou hana i ke kaʻina hana o ka genome decay he mea manaʻo.ʻO ka mea nui, ʻo ke ʻano o ka E. cuniculi ribosome e hōʻike ana i ka nalowale ʻana o ka rRNA a me nā ʻāpana protein i loko o ka microsporidia ribosome e hana i ke kaomi evolutionary e hāpai ana i nā loli i ka ʻano ribosome.Loaʻa kēia mau ʻano like ʻole mai ke kahua ikaika o ka ribosome a ʻike ʻia e kōkua i ka mālama ʻana (a i ʻole e hoʻihoʻi) i ka hui ribosome maikaʻi loa e hoʻopau ʻia e ka rRNA hoʻemi.Hōʻike kēia i kahi mea hou nui o ka microsporidia ribosome i ʻike ʻia i mea e pono ai ke pale i ka hoʻoheheʻe gene.
Ua hōʻike maikaʻi ʻia paha kēia e ka hoʻopaʻa ʻana o ka nucleotide, ʻaʻole i ʻike ʻia ma nā mea ola ʻē aʻe i kēia manawa.ʻO ka ʻoiaʻiʻo o nā koena hoʻopaʻa nucleotide i loko o ka microsporidia maʻamau, akā ʻaʻole i loko o nā eukaryotes ʻē aʻe, e hōʻike ana ʻaʻole nā ​​relics paʻa nucleotide wale nō e kali ana e nalowale, a i ʻole ka pae hope no rRNA e hoʻihoʻi ʻia i ke ʻano o nā nucleotides pākahi.Akā, ʻike ʻia kēia pūnaewele me kahi hiʻohiʻona pono i hiki ke hoʻololi ʻia ma luna o kekahi mau pōʻai koho maikaʻi.ʻO nā kahua hoʻopaʻa nucleotide paha he hopena o ke koho kūlohelohe: ke hoʻohaʻahaʻa ʻia ka ES39L, koi ʻia ka microsporidia e ʻimi i ka uku no ka hoʻihoʻi ʻana i ka biogenesis ribosome maikaʻi loa me ka loaʻa ʻole o ES39L.No ka mea hiki i kēia nucleotide ke hoʻohālike i nā pilina molekala o ka nucleotide A3186 ma ES39L, ua lilo ka nucleotide i poloka hale o ka ribosome, a ua hoʻomaikaʻi hou ʻia ka hoʻopaʻa ʻana ma o ka hoʻololi ʻana o ke kaʻina eL30.
E pili ana i ka molecular evolution o intracellular parasites, hōʻike kā mākou noiʻi ʻana ʻaʻole hana like nā mana o Darwinian koho kūlohelohe a me ka genetic drift o ka genome decay, akā oscillate.ʻO ka mua, hoʻopau ka genetic drift i nā hiʻohiʻona koʻikoʻi o nā biomolecules, e koi nui ana i ka uku.I ka wā wale nō e hoʻokō ai nā parasites i kēia pono ma o ke koho kūlohelohe Darwinian e loaʻa i kā lākou macromolecules ka manawa e hoʻomohala i kā lākou mau hiʻohiʻona maikaʻi loa.ʻO ka mea nui, ʻo ka hoʻololi ʻana o nā kahua paʻa nucleotide i loko o ka ribosome E. cuniculi e hōʻike ana ʻaʻole i hoʻokaʻawale kēia ʻano lilo-i-loaʻa o ka molecular evolution wale nō i ka amortizes deleterious mutations, akā i kekahi manawa hāʻawi i nā hana hou loa i nā macromolecules parasitic.
Ua kūlike kēia manaʻo me ka manaʻo hoʻoneʻe kaulike a Sewell Wright, e ʻōlelo ana he ʻōnaehana koʻikoʻi o ke koho kūlohelohe e kaupalena i ka hiki o nā mea ola ke hana hou51,52,53.Eia nō naʻe, inā hoʻopilikia ka genetic drift i ke koho kūlohelohe, hiki i kēia mau drift ke hana i nā loli i kūpono ʻole iā lākou iho (a i ʻole ka pōʻino) akā alakaʻi i nā hoʻololi hou e hāʻawi i ka hoʻoikaika kino a i ʻole ka hana olaola hou.Kākoʻo kā mākou papa hana i kēia manaʻo ma ka hōʻike ʻana i ke ʻano o ka hoʻololi ʻana e hōʻemi ana i ka paʻi a me ka hana o kahi biomolecule ʻike ʻia ʻo ia ke kumu nui no ka hoʻomaikaʻi ʻana.Ma ka laina me ka win-win evolutionary model, hōʻike kā mākou haʻawina ʻo ka genome decay, i ʻike ʻia ma ke ʻano he kaʻina degenerative, he mea hoʻokele nui hoʻi ia o ka hana hou, i kekahi manawa a i ʻole e ʻae pinepine i nā macromolecules e loaʻa i nā hana parasitic hou.hiki ke hoʻohana iā lākou.