Puas yog Lub Koom Haum Ntawm Qhov Tsis tshua muaj Rs35667974 IFIH1 Gene Polymorphism Nrog Cov Kab Mob autoimmune Ib Tus Mob Ntawm RNA Epigenetics?

Kev pom zoo-AA, AP, thiab EEE, kev saib xyuas-EEE, thiab kev nrhiav nyiaj txiag-EEE. Txhua tus kws sau ntawv tau nyeem thiab pom zoo rau cov ntawv luam tawm ntawm cov ntawv sau. Abstract

Interferon induced nrog helicase C domain-muaj protein ntau 1 (IFIH1) gene encodes cytoplasmic RNA helicase lwm yam hu ua melanoma differentiation-associated 5 (MDA5), RIG-1-zoo li RNA helicase uas paub txog tus kab mob RNA thiab koom nrog innate kev tiv thaiv los ntawm kev paub txog tus kab mob RNA. Raws li kev khi rau ob-stranded (ds) RNA, MDA5 tsim cov filamentous sib dhos raws li qhov ntev ntawm dsRNA thiab siv cov cim kos npe rau kev ntxub ntxaug tus kheej, piv rau tsis yog tus kheej raws li dsRNA ntev thiab methylation. Nws qhov kev hloov pauv tsis zoo rs35667974 yog tiv thaiv hom 1 mob ntshav qab zib, psoriasis, thiab psoriatic mob caj dab, tab sis kuj pom tias muaj feem cuam tshuam nrog kev pheej hmoo siab rau ankylosing spondylitis, Crohn's disease, thiab ulcerative colitis. Txhawm rau kom nkag siab txog lub luag haujlwm nyuaj ntawm qhov sib txawv no peb tau ua qhov kev soj ntsuam ntawm MDA5 hauv txoj hauv kev nrog dsRNA siv molecular dynamics simulations.

Peb cov ntaub ntawv qhia tias thaum Ile923Val kev hloov pauv ntawm rs35667974 variant tsis cuam tshuam rau kev khi rau cov haiv neeg dsRNA loj, nws qhia txog qhov tsis txaus ntseeg nyob rau hauv muaj 2'-O uridine methylation. Yog li, lub xub ntiag ntawm 2′-O-methylation ntawm dsRNA qhia txog kev paub kos npe uas ua rau kev xaiv txo qis ntawm tag nrho MDA catalytic kev ua haujlwm. Txoj kev tshawb no sawv cev rau qhov kev ntsuam xyuas ntawm lub luag haujlwm ntawm kev sib koom rs35667974 variant ntawm autoimmune locus IFIH1, tau tshaj tawm los ua kom txo qis catalytic kev ua haujlwm ntawm kev hloov kho MDA5 phenotype thiab, raws li qhov tshwm sim, txo qis cov lus pom tsis zoo ntawm cytokine thiab chemokine signaling thiab xaiv kev tiv thaiv tiv thaiv autoi. .

Lub helicase C domain yog ib qho tseem ceeb enzyme protein uas muaj peev xwm unwind DNA ob helix qauv. Nws tuaj yeem pab DNA ua kom tiav qhov tseeb replication, kho, thiab xa hauv cov txheej txheem ntawm cell replication, kho, thiab transcription, thiab yog ib tug ntawm cov yuam sij rau lub cev muaj nuj nqi ntawm cell. Nyob rau tib lub sijhawm, kev tiv thaiv kab mob yog ib qho kev tiv thaiv tseem ceeb hauv tib neeg lub cev, uas tuaj yeem tiv thaiv peb ntawm cov kab mob xws li kab mob thiab kab mob.

Cov kev tshawb fawb tau pom tias helicase C domain plays lub luag haujlwm tseem ceeb hauv kev tiv thaiv kab mob. Ua ntej ntawm tag nrho cov, lub helicase C domain tuaj yeem ua kom muaj kev ruaj ntseg thiab coding efficiency ntawm cov noob los ntawm kev pab lub qub replication, kho, thiab transcription ntawm cellular DNA, yog li txhim kho kev tiv thaiv ntawm tib neeg lub cev. Qhov thib ob, lub helicase C domain tuaj yeem txhawb kev paub txog cov protein heterogeneity molecules los ntawm cov hlwb, thiab tswj cov teeb liab hloov ntawm lub cev tiv thaiv kab mob, yog li txhawb lub cev tiv thaiv kev ua haujlwm. Ob qho tib si saum toj no ob txoj hauv kev tau ua lub luag haujlwm zoo hauv kev txhawb nqa kev saib xyuas thiab txhim kho kev tiv thaiv.

Tsis tas li ntawd, qhov kev tshawb fawb tob tob txog kev sib raug zoo ntawm helicase C domain thiab kev tiv thaiv kab mob kuj tseem muab kev tshoov siab tseem ceeb rau peb kom pom cov tshuaj tshiab rau kev kho mob qog noj ntshav thiab kab mob tiv thaiv kab mob. Kev txhim kho tshuaj thiab kev kho noob rau lub hom phiaj helicase C domain tuaj yeem txhim kho lub cev tiv thaiv kab mob qog noj ntshav thiab lwm yam kab mob tiv thaiv kab mob, thiab pab cov neeg mob zoo dua nrog kev kho mob thiab rov zoo ntawm cov kab mob.

Raws li kev txheeb cais, kev sib raug zoo ntawm helicase C domain thiab kev tiv thaiv kab mob yog ze heev, thiab nws ua lub luag haujlwm tseem ceeb hauv kev ua kom muaj kev ruaj ntseg ntawm cov noob, txhawb kev tiv thaiv cellular, thiab txhawb kev tiv thaiv lub cev. Nws vam tias los ntawm cov kev tshawb fawb thiab kev kho mob, peb tuaj yeem tswj hwm thiab txhim kho tib neeg kev tiv thaiv thiab tsim kom muaj kev noj qab haus huv thiab lub neej zoo rau peb. Los ntawm qhov kev xav no, peb yuav tsum txhim kho kev tiv thaiv tus kheej. Cistanche muaj txiaj ntsig zoo rau kev txhim kho kev tiv thaiv vim cov nqaij muab tshuaj txhuam yog nplua nuj nyob rau hauv ntau yam tshuaj antioxidant, xws li vitamin C, vitamin C, carotenoids, thiab lwm yam. Cov khoom xyaw no tuaj yeem tshem tawm cov dawb radicals thiab txo cov kev ntxhov siab oxidative. Txhawb nqa thiab txhim kho kev tiv thaiv ntawm lub cev tiv thaiv kab mob.

Nyem cistanche tubulosa cov txiaj ntsig

Ntsiab lus

Single-nucleotide polymorphism (SNP) · Molecular model · Interferon induced with helicase C domain 1 (IFIH1) · Melanoma differentiation-associated 5 (MDA5) · RNA methylation.

Taw qhia

Cov noob thiab cov txheej txheem koom nrog cov kab mob autoimmune, uas cuam tshuam txog kwv yees li 5 feem pua ​​​​ntawm cov pej xeem, tseem tsis paub tab sis khaws cov ntaub ntawv xav tau tias cov kab mob sib txawv ntawm cov kab mob autoimmune tuaj yeem qhia txog keeb kwm ntawm caj ces, yog li taw qhia txog qhov muaj ntau yam sib txawv uas sib koom los ntawm cov kab mob autoimmune sib txawv. (Zhernakova et al. 2009). Kev sim tshem tawm cov ntaub ntawv caj ces no rau hauv cov txheej txheem lom neeg muaj txiaj ntsig ua rau cov kab mob hais txog kev txheeb xyuas cov noob caj noob ces. Kev txheeb xyuas cov kab mob sib txawv yog ib txoj haujlwm nyuaj tab sis tsim nyog hauv kev sim tsim txoj hauv kev zoo rau kev kwv yees kab mob, kev tiv thaiv, thiab kev cuam tshuam (Biros et al. 2005).

Ntau hom RNA molecules koom nrog hauv kev tswj hwm ntawm ntau cov txheej txheem lom neeg, suav nrog tus xa xov RNA (mRNA), hloov RNA (tRNA), ribosomal RNA (rRNA), microRNA (miRNA), thiab ntev noncoding RNA (lncRNA). RNA molecules muaj ntau (ntau tshaj 150) tshuaj hloov kho (Machnicka li al. 2013; Boccaletto li al. 2018). Cov kev hloov kho no tau ua haujlwm txuas mus rau txhua theem ntawm RNA metabolism, xws li cov qauv, kev ruaj ntseg, thiab kev sib cuam tshuam, thiab ua lub luag haujlwm tseem ceeb hauv ntau cov txheej txheem lom neeg, xws li kev hloov pauv ntawm cov kab mob thiab cov tshuaj tiv thaiv kab mob (Machnicka li al. 2013). Ntawm lawv, ribose methylation yog ib qho kev hloov kho uas muaj nyob hauv RNA. 2′-O -methyluridine muaj nyob rau hauv rRNA, snRNA, snoRNA, thiab tRNA ntawm Archaea, Bacteria, thiab Eukaryota (Aučynaitė li al. 2018). Lub ribose-2'-Omethylation nce hydrophobicity ntawm nucleotides thiab tiv thaiv lawv tawm tsam kev ua ntawm nucleases (Yildirim li al. 2014).
Cov ntaub ntawv pov thawj qhia tau hais tias 2′-O-methylation ntawm tus kab mob RNA (2′Ome-RNA) ua lub luag haujlwm tseem ceeb hauv kev khiav tawm ntawm lub cev tiv thaiv kab mob hauv lub cev hauv lub cev (Dimitrova li al. 2019). Züst thiab cov npoj yaig tau qhia tias 2′ OMe ntawm tus kab mob RNA tau pab txhawb rau kev khiav tawm ntawm interferon (IFN)-cov tshuaj tiv thaiv kab mob sib kis, yog li txhawb nqa cov kab mob sib kis (Züst li al. 2011). Tsis tas li ntawd, Vitali thiab Scadden tau hais tias IU-dsDNA suppresses MDA5 IFN stimulating txoj kev (Vitali thiab Scadden 2010).

Interferon induced nrog helicase C domain 1 (IFIH1) gene encodes cytoplasmic RNA helicase tseem hu ua MDA5 (Melanoma differentiation-associated protein 5), thiab nws yog RIG-I-zoo li receptor (RLR) uas ua haujlwm tiv thaiv kab mob hauv lub cev tiv thaiv kab mob. los ntawm kev kuaj pom tus kab mob RNAs. MDA5 lees paub 0.5–1 kb RNA duplex qia qauv uas feem ntau yog tsim thaum lub sij hawm picornaviral replication thiab kho kom haum rau lub cev tiv thaiv kab mob (Nejenttsev li al. 2009; Crow 2011). MDA5, thaum tshawb pom cov kab mob ntev ntev ob-stranded RNAs (dsRNAs), tsim tawm thaum lub sij hawm rov ua dua ntawm picornaviruses, qhib hom I interferon signaling txoj hauv kev. Cov kev tshawb fawb tau pom tias MDA5 tsim cov filament raws qhov ntev ntawm dsRNA thiab siv ATP-dependent filament dynamics rau kev ntxub ntxaug ntawm tus kheej thiab tsis yog tus kheej raws li dsRNA ntev (Toro li al. 2015). MDA5 tau pom tias muaj kev koom tes hauv kev hloov pauv ntawm crosstalk ntawm cov hlwb thiab lub cev tsis muaj zog / hloov kho lub cev los ntawm kev tsim khoom hauv zos ntawm cytokines thiab chemokines.

Kev hloov pauv hauv MDA5 kev qhia thiab/lossis kev ua ub no yuav ua rau cov lus teb ntawm tes rau dsRNA, ib qho khoom los ntawm tus kab mob replication (Colli et al. 2010). Nws kuj tau pom tias kev hloov pauv ntawm flament forming-associated residues ua rau poob ntawm filament formation thiab MDA5- nyob ntawm kev taw qhia, tshwj tsis yog rau ib khub ntawm kev hloov pauv, uas txhim kho cov teeb liab. Cov txiaj ntsig no tau qhia tias ATP-ywj pheej ntawm cov txheej txheem, piv txwv li, nruj dua RNA khi thiab / lossis ruaj khov ntawm cov protein-protein kev sib cuam tshuam, zoo li yuav yog lub luag haujlwm rau kev ruaj ntseg ntawm MDA5 filament tsim hauv vitro thiab rau kev ua haujlwm siab dua hauv cov hlwb (Sohn thiab Xyoo 2016).

Smyth et al. (2006) and Nejentsev et al. (2009) tau piav qhia txog qhov tsis tshua muaj allele ntawm IFIH1 noob uas muaj kev tiv thaiv los ntawm hom 1 ntshav qab zib (T1D). Qhov no rs35667974 IFIH1 ib leeg-nucleotide polymorphism (SNP), qhov chaw khaws cia isoleucine (codon [ATT]) ntawm txoj hauj lwm #923 hloov mus rau valine (codon [GTT]), yog qhov tsis tshua muaj qhov sib txawv raws li qhov me me allele zaus (MAF) yog C {{ 9}}.010031 (2655 tus neeg hauv tag nrho cov qauv ntawm 264690) raws li TOPMED (Taliun et al. 2021) thiab C = 0.016267 (3343 tus neeg hauv tag nrho cov qauv ntawm 205514) raws li ALFA (Phan et al. 2020; Sherry et al. 2001). Hauv Table 1 tib neeg biogeography ntawm zaus ntawm polymorphism nyob rau hauv txawv teb chaws cheeb tsam raws li ALFA project data (Phan li al. 2020; Sherry li al. 2001), yog nthuav tawm. Cov kev tshawb fawb tom qab tau lees paub tias qhov tsis tshua muaj allele no muaj tib yam cuam tshuam rau T1D, psoriasis (PS) (Li et al. 2010), thiab psoriatic mob caj dab (PsA) (Budu-Aggrey li al. 2017). Ntawm qhov tsis sib xws, SNP no tau cuam tshuam los ntawm qhov muaj feem cuam tshuam rau kev cuam tshuam ntawm kev txhim kho ankylosing spondylitis (AS) (Ellinghaus li al. 2016), Crohn's disease (CD) (Ellinghaus et al. 2016; Budu-Aggrey li al. 2017). ), thiab ulcerative colitis (UC) (Ellinghaus li al. 2016; Budu-Aggrey li al. 2017).

Chistiakov et al. (2010) tau qhia tias poob-ntawm-function mutations E627X thiab I923V ntawm MDA5 yog txuam nrog qis poly(I: C)-induced interferon- tsim nyob rau hauv peripheral ntshav mononuclear hlwb ntawm hom 1 mob ntshav qab zib thiab yog li ntawd yog T1D tiv thaiv. Nws tseem tau hais tias nyob rau hauv MDA5 molecule, I923V amino acid hloov pauv nyob rau thaj tsam ntawm H927 residue, uas pab txhawb rau kev khi ntawm dsRNA (Yoneyama thiab Fujita 2008). Txawm li cas los xij, MDA variant I923V tau pom tias muaj peev xwm los khi dsRNA tab sis ib qho 2.5-fold txo cov kev ua haujlwm catalytic (Shigemoto li al. 2009). Yog li ntawd, qhov polymorphism no zoo li tsis cuam tshuam rau txoj hauv kev tseem ceeb ntawm nucleotide acid-binding zog ntawm no cytoplasmic RNA-sensing helicase tab sis hloov nws txoj haujlwm los ntawm ib qho tseem tsis tau paub txog.

Kev sib raug zoo ntawm IFHI1 polymorphism thiab qhov tshwm sim ntawm tus kab mob enterovirus hauv T1D thiab kev koom tes ntawm MDA5 I923V variant thiab zaus ntawm enteroviral RNA hauv cov neeg mob T1D tau pom (Looney li al. 2015). Tsis tas li ntawd, cov kev tshawb fawb tsis ntev los no ntawm MDA5- thiab MAVSknockout nas tau pom lub luag haujlwm tseem ceeb ntawm cov proteins no hauv kev sib kho hom 1 interferon cov lus teb tawm tsam tus kab mob Coxsackie B (Wang et al. 2010). Ib tug mengovirus tus thawj coj protein tau pom los tiv thaiv kev qhia ntawm IFN- los ntawm kev thaiv cov dimerization ntawm IRF3 xav tau rau kev ua kom muaj qhov cuam tshuam no (Hato li al. 2007). Qhov kev soj ntsuam no qhia tias cov kev hloov pauv uas cuam tshuam IFIH1 kev ua haujlwm hauv lub tswv yim tshuaj tiv thaiv kab mob tau raug xaiv tsis zoo, tsis yog xaiv qhov zoo vim tias lawv tau txais kev tiv thaiv los ntawm T1D (Crow 2011).

Chaw et al. (2018) tau txheeb xyuas ntau yam ntawm lwm tus RIG-I-zoo li receptors, thaum Brisse thiab Ly (2019) tau tshuaj xyuas ntau qhov kev hloov pauv thiab qhov tshwj xeeb ntawm MDA5 thiab nws qhov cuam tshuam RIG-I. Qhov chaw ntawm cov hloov pauv residues nyob rau hauv los yog ze ntawm RNA-binding thiab ATP-binding qhov chaw los yog cov filament interface coj peb mus rau hypothesize hais tias cov kev hloov pauv tau pom yuav txhim khu kev ruaj ntseg ntawm IFIH1 filament los ntawm kev ua kom cov intrinsic affinity ntawm IFIH1 thiab dsRNA los yog ntawm IFIH1. molecules nyob rau hauv filament los yog los ntawm kev txo cov efficiency ntawm ATP hydrolysis thiab, yog li, filament disassembly tus nqi (Rice li al. 2014).

Qhov kev ua haujlwm no sawv cev rau kev tshawb fawb txog kev muaj peev xwm ntawm lub luag haujlwm ntawm kev sib koom rs35667974 variant ntawm autoimmune locus IFIH1, tau tshaj tawm ua rau muaj kev cuam tshuam rau kev ua haujlwm phenotype encoding Ile923Val amino acid hloov pauv hauv IFIH1 gene protein khoom MDA5. Cov tom kawg yog ib qho kev sib tw lom neeg uas muaj feem cuam tshuam rau cov noob caj noob ces sib koom nrog ntau yam kab mob, cuam tshuam rau kev tswj hwm hauv zos ntawm cytokines thiab chemokines uas tiv thaiv autoimmunity (Colli li al. 2010). Qhov kev ua haujlwm no yog tsom mus tshawb txog qhov tseem tsis tau paub txog qhov kev hloov pauv ntawm Ile923Val txo cov kev ua haujlwm catalytic ntawm tib neeg MDA5. Hauv txoj kev tshawb no, peb tau tshawb xyuas qhov sib txawv ntawm kev sib cuam tshuam ntawm MDA5 nrog dsRNA ntawm haiv neeg thiab qhov tsis tshua muaj qhov txawv ntawm Ile923Val hauv kev pib ntawm cov txheej txheem o. Raws li, peb tsom mus soj ntsuam tus cwj pwm zoo ntawm tib neeg MDA5 / dsRNA complex nyob rau hauv ib qho chaw aqueous nyob rau hauv lub xub ntiag ntawm Ile923 los yog Val923 thaum uracil 2′-O yog methylated los yog tsis. Qhov kev soj ntsuam ntawm cov qauv ntawm cov caj ces tsis tshua muaj kev cuam tshuam los yog kev tiv thaiv loci tuaj yeem muab kev nkag siab rau peb txoj kev nkag siab ntawm pathophysiology ntawm cov kab mob autoimmune thiab cov kev tshawb fawb tshawb pom yuav cuam tshuam rau kev tswj xyuas cov kab mob zoo dua hauv kev kawm.

Cov ntaub ntawv thiab cov txheej txheem

Kev Tshawb Fawb Txog Kev Tshawb Fawb, Kev Tsim Kho Ntoo Phylogenetic, thiab Kev Tshawb Fawb Kev Xaiv Zoo

Cov protein ntau ntawm Homo sapiens (ib ntu ID: NP_071451.2) tau muab los ntawm UniProt database (The UniProt Consortium 2021). Txhawm rau nrhiav homologs thoob plaws hom, BLAST tshawb nrhiav tau ua nrog Mega BLAST (National Center for Biotechnology Information, NCBI, Bethesda, MD, USA) ntawm RefSeq thiab NR protein database (thiab PDB thiab UniProt) siv Blastp (protein-protein BLAST) nrog default parameters (Altschul li al. 1997). 1000 homologs rau tib neeg MDA5 cov proteins tau raug xaiv thawj zaug thiab kev xaiv thoob plaws hom tsom rau C-terminal domain uas muaj cov kab ke ib puag ncig tib neeg I923V hloov pauv tau siv los txheeb xyuas qhov kev hloov pauv no hauv lwm hom. Clustal Omega, ntau qhov kev sib txuas ua ke (Clustal-O) (Sievers li al. 2011), thiab T-Cofee ntau qhov sib txuas ua ke server (Notredame li al. 2000; Di Tommaso li al. 2011) tau siv los ua cov protein ntau sib npaug. thiab Unipro UGENE platform bioinformatics software (Okonechnikov et al. 2012) los xaiv pom ntau yam kev sib dhos.

Kev tsom xam evolutionary yog siv los txheeb xyuas cov haujlwm ntawm cov protein ntau uas tau txais kev txuag ntau ntawm cov tsiaj, qhia txog qhov tseem ceeb ntawm cov qauv (Andreou li al. 2018). Cov ntoo phylogenetic tau tsim los siv txoj kev Tshaj Lij Tshaj Lij Tshaj Lij (Nei thiab Kumar 2000) thiab tus qauv Tamura-Nei (Tamura thiab Nei 1993) nrog 500 bootstrap replicates (Felsenstein 1985). Thawj tsob ntoo (s) rau qhov kev tshawb nrhiav heuristic tau txais los ntawm kev thov Neighbor-Join thiab BioNJ algorithms rau cov matrix ntawm kev sib txuas sib txuas kwv yees siv tus qauv Tamura-Nei thiab tom qab ntawd xaiv cov topology nrog cov txiaj ntsig zoo tshaj plaws. Kev tshuaj xyuas phylogenetic koom nrog 52 homolog nucleotide sequences (39 orthologs thiab 13 paralogs) ntawm tib neeg IFIH1 noob. Codon txoj haujlwm suav nrog yog 1st ntxiv rau 2nd ntxiv rau 3rd ntxiv rau Noncoding. Muaj tag nrho ntawm 3729 txoj haujlwm hauv cov ntaub ntawv kawg. Kev tshuaj ntsuam Evolutionary tau ua los ntawm MEGA11 software pob (Tamura li al. 2021).

Txhawm rau txheeb xyuas seb IFIH1 noob tau hloov pauv hloov pauv li cas, peb tau siv qhov program codeml hauv PAML v4.9j software pob (Yang 2007). Cov kab ke nucleotide thiab cov ntaub ntawv sib txuas ntawm cov protein sib txuas tau raug xa mus rau PAL2NAL (Suyama li al. 2006) los tsim cov ntaub ntawv tsim nyog CODEML input nucleotide alignment files. Qhov kev xaiv zoo tshuaj ntsuam xyuas rau ortholog MDA5 cov noob tau ua los ntawm cov qauv ntawm qhov chaw thiab cov ceg ntoo (Yang li al. 2005; Yang and Bielawski 2000). Qhov tsis sib xws / sib piv tus nqi hloov pauv (ω=dN / dS) muab qhov ntsuas ntawm kev xaiv siab ntawm qib amino acid. Qhov loj ntawm tus nqi ntawm dN / dS (ω) sawv cev rau hom kev xaiv: ω<1 for negative selection, ω=1 for neutral selection, and ω>1 rau kev xaiv zoo (Yang et al. 2005). Hauv CODEML cov qauv ntawm qhov chaw (M0, M1, M2, M3, M7, thiab M8) thiab ceg-site qauv (Clades A thiab C) raug xaiv los ua qhov kev xaiv zoo (Bielawski thiab Yang 2004; Yang thiab Nielsen 2002). Hauv cov qauv ntawm qhov chaw, qhov kev xeem qhov piv txwv (LRT) tau siv los ntsuas qhov kev xaiv zoo los ntawm kev sib piv peb khub ntawm cov qauv (M0 / M3, M2 / M1, thiab M7 / M8). Kev tsom xam tau ua ob qho tib si rau qhov ntev-ntev ib ntus thiab C-Terminal Domain (CTD) ib ntus.

Kev Ntsuam Xyuas Cov Qauv thiab Molecular Dynamics Simulations

Cov qauv cryoelectron microscopy (cryo-EM) qauv ntawm hMDA5-dsRNA filament nyob rau hauv lub xub ntiag ntawm ATP (PDB ID: 6GKM) (Yu li al. 2018) (Berman li al. 2000) tau siv los ua ib tug qauv system rau lub molecular dynamics ( MD) simulations. Tag nrho cov protein residues daws tau (307–1020), 14 lub hauv paus khub ntawm ob-stranded RNA (dsRNA), thiab cov zinc sib koom ua ke tau khaws cia, qhov seem uas ploj lawm tau ua qauv siv SWISS-MODEL server (Waterhouse li al. 2018). Force field parameters thiab hydrogen atoms tau ntxiv siv XLEaP module ntawm AMBER 18 (Case et al. 2005). AMBER force fields f14SB (Maier et al. 2015) thiab f99OL3 (Zgarbová et al. 2011) tau siv rau cov protein thiab RNA, feem, nrog rau cov modrna08 (Aduri et al. 2007) parameters for modified nucleosides. I923V kev hloov pauv tau qhia hauv MDA5 los ntawm kev tshem tawm Cδ methyl pawg ntawm I923, thaum hloov kho ribose ntawm U12 yog methylated ntawm 2′-O siv lub tsev qiv ntawv modrna08 residue MRU. Cov zinc ion tau sib txuas nrog 4 cysteine ​​residues 907, 910, 962, thiab 964, siv cov kev quab yuam kom tsim nyog los tuav lub tetrahedral coordination sphere (Zn-S daim ntawv cog lus ntev ntawm 2.35 Å nrog 50 kcal·mol–1·Å–2 quab yuam tsis tu ncua thiab S-Zn-S kaum ntawm 109.5 deg nrog 25 kcal·mol–1·rad–2).

Ua li no, peb npaj 4 lub tshuab rau MD simulations: (i) haiv neeg MDA5–dsRNA, (ii) MDA5(V923)–dsRNA, (iii) MDA5–dsRNA(2′Ome), thiab (iv) MDA5( V923)–dsRNA(2′OMe). Tag nrho cov tshuab tau raug daws nyob rau hauv truncated octahedral hnyav thawv ntawm pre-equilibrated TIP3P dej molecules, nrog ib tug yam tsawg kawg nkaus tsis ntawm 10 Å nyob ib ncig ntawm lub complex thiab ces tus yuav tsum tau tus naj npawb ntawm counter ions tau ntxiv kom tau tus nqi neutralization ntawm lub tshuab. MD simulations tau ua nrog GPU-accelerated version ntawm PMEMD (Salomon-Ferrer li al. 2013) module hauv AMBER 18 thiab lub sijhawm kauj ruam ntawm 2 fs. Qhov ntsuas kub tau tswj hwm siv Langevin thermostat nrog kev sib tsoo ntau zaus ntawm 1.0 ps-1, thaum lub siab tswj tau siv Berendsen barostat nrog lub sijhawm so siab ntawm 1.0 ps. SHAKE tau siv los txwv cov ntawv cog lus nrog hydrogen atoms nrog kev ua siab ntev ntawm 10-6 Å, qhov tsis muaj kev sib cuam tshuam tau suav nrog qhov chaw ncaj qha ntawm 10 Å.

Kev txo qis zog tau ua tiav thawj zaug rau 1{{20}},000 cov kauj ruam nrog rau qhov chaw txwv ntawm 100 kcal·mol–1·Å–2 quab yuam tas li ntawm cov tsis-hydrogen atoms ntawm MDA5-dsRNA. Cov kuab tshuaj tau sib npaug ntawm 300 K thiab 1 atm los ntawm kev sib tw luv luv ntawm kev sim hauv NVT thiab NPT pawg, 100 ps thiab 400 ps, ​​raws li, thaum tuav cov kev txwv ntawm cov tsis-hydrogen atoms ntawm solute. Tom qab ntawd, lub zog txo qis tau ua rau 10,{13}} cov kauj ruam, tab sis nrog kev txwv tsis pub muaj 10 kcal·mol–1·Å–2 tsuas yog ntawm C atoms ntawm MDA5 thiab phosphate caj qaum ntawm dsRNA. Nyob rau hauv 3 sub tau maj mam so (10.0, 1.0, 0.1 kcal·mol–1·Å–2) thoob plaws 1 ns, ua raws li 9 ns ntawm unrestraint equilibration nyob rau hauv tas li lub siab. Tom qab cov thawj 10 ns ntawm kev sib npaug (tsis siv hauv kev tshuaj ntsuam), 100 ns ntawm kev tsim khoom simulations hauv NPT pawg tau ua rau txhua qhov system ntawm 300 K thiab 1 atm, thaum khaws cov snapshots ntawm lub kaw lus txhua 5.0 ps rau kev tsom xam siv CPPTRAJ module ntawm AMBER 18 (Roe thiab Cheatham 2013). Txhua daim duab piav qhia 3D qauv tau tsim los siv PyMOL molecular graphics system (v.2.3 qhib-qhov tsim).

Cov txiaj ntsig

Phylogenetic Analysis ntawm Ile923Val Hloov ntawm MDA5

Qhov kev hloov pauv thiab qhov tshwj xeeb ntawm MDA5 thiab nws qhov cuam tshuam RIG-I tau raug tshuaj xyuas los ntawm (Brisse thiab Ly 2019). Ntawm no IFIH1 evolution tau ua haujlwm los txhais cov ntsiab lus kev txuag hauv MDA5 ib ntus txog cov polymorphism tshwj xeeb. Kev tsom xam Evolutionary tau qhia txog kev txuag hnyav ntawm MDA5 ntawm ntau hom (984 ntawm 1000 qhov kev tshawb nrhiav muaj isoleucine ntawm qhov sib npaug ntawm hMDA # 923) hauv RD / CTD domain, qhia txog cov qauv / kev ua haujlwm tseem ceeb. Lub polymorphism rs35667974 hauv exon 14 ntawm tib neeg cov noob ua rau muaj kev txuag amino acid hloov ntawm txoj hauj lwm 923 los ntawm Ile mus rau Val hauv hMDA5. Txawm li cas los xij, muaj lwm kaum rau hom kab nyob deb uas muaj tib txoj hauj lwm nyob hauv valine (Daim duab 1) qhia txog kev muaj peev xwm ntawm qhov kev hloov no hla cov tsiaj hauv RD/CTD domain ntawm MDA5 thiab qhov ze ze homologous RS / GY domain ntawm isoforms X1, X3, thiab DHX58 helicases (Fig. 2). Tsis tas li ntawd, kev sib txuas ua ke ntawm thaj av ib ncig ntawm hMDA # 923 txoj haujlwm (MDAs CTD kev sib cuam tshuam lub voj voog) qhia txog qhov nruab nrab rau kev txuag tau zoo ntawm cov tsiaj nyob deb uas qhia txog thoob plaws hom kev ua haujlwm tseem ceeb hauv cheeb tsam.

Kev Tshawb Nrhiav Kev Xaiv Zoo

Txhawm rau txheeb xyuas seb IFIH1 noob tau hloov pauv hloov pauv ntawm qhov chaw ua qauv thiab cov qauv ntawm cov ceg ntoo tau siv los ua qhov kev xaiv zoo ntawm orthologs ntawm tag nrho cov noob thiab CTD sau. Hauv cov qauv ntawm qhov chaw, tsis muaj qhov chaw xaiv zoo tau txheeb xyuas rau CTD sau (Table 2). M0 txhais tau tias qhov kev hloov pauv tsis tu ncua (ω=dN/dS=0.2) (Table 2). Qee qhov chaw uas tau txais kev xaiv zoo tau raug txheeb xyuas siv M2- thiab M8-site qauv qauv rau tag nrho cov noob (Suppl. Table 1), txawm hais tias tsis nyob rau hauv RNA interacting regions thiab CTD domain. Hauv cov qauv ntawm qhov chaw, ω (dN / dS) yog<1 which indicates a highly conserved gene (Table 2, Suppl. Table 1). In the branch-site model, the human branch (as well as the primate's branch) was used as the foreground clade, the ω value was low, and no sites with posterior probability greater than 0.85 were identified (Suppl. Table 2).

Tshwj xeeb, rau cov ceg-site qauv C rau CTD sau (Suppl. Table 3), 33 feem pua ​​​​ntawm cov chaw tau hloov zuj zus ntawm qeb ω0=0. 036. Raws li cov chaw hloov kho nyob rau hauv pawg no tsis paub qhov txawv ntawm hom ceg, ob hom ceg muaj nqi tib yam ntawm ω rau cov chaw hauv pawg no. Ntxiv mus, 55 feem pua ​​​​ntawm cov vev xaib tau hloov zuj zus raws li qeb ω2. Txawm li cas los xij, cov no muaj ω qhov tseem ceeb uas yog nyob ntawm hom ceg (ω20=0.25 thiab ω21=0). Tsis tas li ntawd, rau ceg-site qauv C rau 39 ortholog MDA5 noob (Suppl. Table 4), 33 feem pua ​​​​ntawm cov chaw tau hloov zuj zus ntawm qeb ω0=0. 027. Ntawm qhov tod tes, 41 feem pua ​​​​ntawm cov vev xaib tau hloov kho raws li qeb ω2. Txawm li cas los xij, cov no muaj ω qhov tseem ceeb uas yog nyob ntawm hom ceg (ω20=0.25 thiab ω21=15.32).

Kev Ntsuam Xyuas Cov Qauv

Qhov kev soj ntsuam evolutionary ua pov thawj tias Ile923Val hloov pauv tsis yog qhov txawv txav ntawm tib neeg hom, txij li Val muaj nyob rau hauv MDA5 txoj haujlwm nyob rau lwm hom ib yam nkaus. MDA5 yog tus kab mob kis tau ob-stranded RNA (dsRNA) receptor uas ua lub luag haujlwm tseem ceeb hauv kev tiv thaiv kab mob los ntawm nws qhov tshwj xeeb rau tus kab mob RNA (Wu li al. 2013). Nws tau raug pom tias 2′-O-methylation ntawm tus kab mob mRNA yog ib qho tseem ceeb rau kev tiv thaiv kab mob hauv lub cev, yog li nws tau pom tias 2′-O-methylation yog molecular kos npe rau qhov sib txawv ntawm tus kheej tiv thaiv tsis yog tus kheej mRNA. (Zust et al. 2011). Txhawm rau tshawb xyuas lub luag haujlwm muaj peev xwm ntawm Ile923Val hloov pauv hauv qhov tsis txaus ntseeg IFIH1 variant rs35667974, peb tau tshuaj xyuas cov qauv cryoelectron microscopy (cryo-EM) ntawm MDA5-dsRNA filament nyob rau hauv lub xub ntiag ntawm ATP (PDB ID: 6GKM) (Yu et al. 2018). Txoj hauj lwm 923 yog nyob rau ntawm lub voj 921-927 uas cuam tshuam ncaj qha nrog dsRNA (Daim duab 3A). Tshwj xeeb, Ile923 yog nyob ntawm 4.8 Å los ntawm 2′-ΟΗ ntawm uridine U12, thiab lawv cov kev sib cuam tshuam yog ruaj khov los ntawm kev sib cog lus hydrogen ntawm qhov uas nyob ib sab His927 thiab uracil puag. Hloov pauv ntawm Ile923 los ntawm Val hauv rs35667974 variant tsis xav tias yuav qhia txog kev sib cav tsis sib haum xeeb, tsis yog txo qis kev cuam tshuam nrog 2′-ΟΗ ntawm uridine U12 (Fig. 3B). Nyob rau hauv rooj plaub RNA yog methylated ntawm ribose ntawm U12, ces lub ntuj MDA5 variant nrog Ile923 qhia ib tug haum van der Waals hu nrog ib tug 2′-OMe pab pawg neeg ntawm U12 ntawm 3.6 Å (Fig. 3C), whereas Val923 ntawm lub rs35667974 variant yog nyob ntawm 5.0 Å (Fig. 3D). Cov kev sib txawv no tsis tuaj yeem qhia qhov cuam tshuam loj ntawm Ile923Val hloov pauv ib se; Txawm li cas los xij, kev hloov pauv hloov maj mam feem ntau ua rau muaj kev hloov pauv tseem ceeb los ntawm kev cuam tshuam ntawm cov qauv kev ua haujlwm ntawm lub cev.

Molecular Dynamics suav

Txhawm rau tshawb xyuas cov txiaj ntsig ntawm I923V kev hloov pauv ntawm MDA5 hauv nws qhov kev cuam tshuam nrog dsRNA, ob qho tib si ib txwm muaj thiab 2′-Ο-methylated, peb tau ua haujlwm molecular dynamics simulations ntawm 4 lub tshuab ntawm 100-ns timescale. Lub zog ntawm cov tshuab tau saib xyuas siv lub hauv paus-lub ntsiab lus-square fluctuations (RMSF) ntawm txhua cov protein residue thiab hydrogen-bonding nrug ntawm H927 nrog U12 (Fig. 4). Peb cov kev suav suav hais tias kev hloov pauv ntawm I923 rau V923 hauv MDA5 ua rau muaj kev cuam tshuam me me ntawm cov dynamics hauv cheeb tsam RNA tiv tauj (residues 923-934) thiab interprotein kev sib tshuam voj (950-955) ntawm cov complex nrog haiv neeg dsRNA (Fig. 4C) . Qhov kev soj ntsuam no zoo ib yam li ntawm 2'-O-methylation hauv U12, txawm hais tias muaj cov txiaj ntsig zoo tshaj plaws tau pom nyob rau hauv tag nrho cov dynamics ntawm thaj tsam carboxy-terminal ntawm MDA5 V923 mutant (Fig. 4D).

Tam sim no xav txog qhov tseem ceeb hydrogen-bonding kev sib cuam tshuam ntawm H927 residue uas nyob ib sab nrog lub hauv paus uracil, peb MD simulations qhia tias methylation ntawm 2′-O tsis cuam tshuam rau nws hauv haiv neeg MDA5 (Fig. 4E). Txawm li cas los xij, qhov kev hloov pauv ntawm V923 tsis cuam tshuam rau hydrogen daim ntawv cog lus ntawm H927 hauv cov haiv neeg dsRNA tab sis pom muaj qhov tsis txaus ntseeg nyob rau hauv muaj 2'-O-methylation (Daim duab 4F). Ua ke, peb MD simulations qhia tias txawm hais tias cov nyhuv ntawm I923V kev hloov ntawm MDA5 nyob rau hauv kev sib cuam tshuam nrog haiv neeg dsRNA yog marginal, nws cov nyhuv nyob rau hauv lub zog thiab stability ntawm lub MDA5 / RNA complex yog tseem ceeb dua thaum uracil yog 2′-O-methylated. .

Kev sib tham

Txoj kev tshawb no sawv cev rau kev tshawb nrhiav evolutionary thiab cov qauv ntawm lub luag haujlwm ntawm kev sib koom rs35667974 variant ntawm autoimmune locus IFIH1, tau tshaj tawm ua rau kev hloov kho phenotype rau MDA5 (Downes li al. 2010). Daim ntawv thov ntawm evolutionary site thiab ceg-site qauv los ua qhov zoo xaiv tsom xam qhia tias tsis muaj qhov zoo xaiv qhov chaw hauv RNA kev sib cuam tshuam qhov chaw uas rs35667974 polymorphism nyob. Tsis yog cov qauv ntawm cov chaw ua haujlwm qhia tias cov chaw tshwj xeeb tau xaiv qhov zoo hauv CTD sau.

Qhov no yog nyob rau hauv kev pom zoo nrog cov tsos mob tsawg heev ntawm Ile923Val variant txawm ntawm cov tib neeg cov pejxeem (Table 1). Txawm li cas los xij, cov neeg nyob sab Europe hauv kev sib piv nrog lwm tus muaj qhov sib txawv ntawm qhov zaus ntawm C allele ntawm ib qho kev txiav txim loj. Yog tsis saib xyuas tus qauv me me, thaj chaw faib khoom ntawm polymorphism nyob rau hauv txoj kev tshawb fawb qhia tau hais tias nws cov tsos nyob rau hauv cov neeg nyob sab Europe raws li qhov pib taw tes yuav yog vim nyob rau hauv tej yam kev mob thiab khoom noj khoom haus uas tau hloov nyob rau hauv xyoo tas los. Kev tshawb nrhiav cov qauv ntawm lub luag haujlwm ntawm kev kuaj xyuas SNP tau ua los ntawm kev tshuaj xyuas cov qauv ntawm dsRNA-MDA5 (native thiab mutant) complex.

Ntawm dsRNA-MDA5 txoj kev tsim theem kev taw qhia ntawm Ile923Val kev hloov pauv cuam tshuam kev cuam tshuam ntawm cov protein C-terminal domain (CTD) nrog dsRNA nrog kev taw qhia ntawm hydrophobic kab noj hniav ntawm ib sab ntawm ribose qab zib ntawm ib-strand ntawm dsRNA. . Peb tau pom tias nyob rau hauv cov ntaub ntawv ntawm methylated RNA ntawm phosphoribosyl saw ntawm ib tug strand, ntxiv dynamic teebmeem yuav cuam tshuam rau kev sib cuam tshuam ntawm cov mutant thaum tsis cuam tshuam rau cov qus hom. Qhov no tau pom zoo nrog cov kev tshawb fawb kev sim (Looney li al. 2015; Brisse thiab Ly 2019) qhia tias cov nyhuv ntawm Ile923Val polymorphism txheeb xyuas nyob rau thaj tsam ntawm MDA5-dsDNA kev sib cuam tshuam taw tes yuav tsis cuam tshuam rau haiv neeg dsRNA-binding zog tab sis hloov los ntawm 2. Tsis tas li ntawd, cov kev tshawb fawb molecular dynamic tau qhia txog lub luag haujlwm tseem ceeb ntawm methylation hauv cov mutant hais txog kev txav mus los thiab kev ruaj ntseg ntawm MDA5 loops 941-959 thiab 970-977 koom nrog dsRNA kev sib cuam tshuam thiab kev sib cuam tshuam hauv MDA filament tsim. Cov teebmeem zoo li no tuaj yeem cuam tshuam rau MDA5 filament assembly, MDA5-MAVS lub koom haum, thiab MAVS filament assembly uas txuas ntxiv qhib qhov kev qhia ntawm hom I interferon noob (IFN1: IFN thiab IFN ). Hauv qhov xwm txheej ntawm T1D thiab PsA, raws li qhov tshwm sim, txo qis ntawm MDA5 cov protein ua haujlwm thiab yog li txo qis IFN ntau lawm tiv thaiv autoimmunity. Cov kev soj ntsuam no qhia tias ntau qhov sib txawv ntawm IFIH1, kwv yees tias yuav cuam tshuam rau kev sib cuam tshuam ntawm MDA5 rau MAVS thiab kev txo qis ntawm IFN ntau lawm yuav txo qhov kev pheej hmoo ntawm cov kab mob, thaum ib txwm ua haujlwm MDA5 cuam tshuam nrog lawv (Shigemoto li al. 2009).

Qhov no ua rau kev txiav txim siab tias ib yam li tus kab mob RNA, tus kheej-dsRNA methylation, kev hloov pauv ntawm RNA, yog qhov tseem ceeb ntawm kev xaiv / ua kom muaj txiaj ntsig rau kev qhia txog kev tiv thaiv hauv qee cov kab mob autoimmune. Cov txiaj ntsig los ntawm kev tshawb fawb tam sim no nthuav dav kev paub txog kev lom neeg tseem ceeb ntawm rs35667974 SNP ntawm IFIH1 locus hauv kev txhim kho cov kab mob uas tau hais tseg thiab qhia txog qhov tseem ceeb ntawm kev tshawb fawb ntawm cov noob sib koom los ntawm ntau yam kab mob autoimmune. Txawm li cas los xij, hauv kev tshawb fawb pej xeem rau kev sib txuas caj ces ntawm SNPs nrog cov kab mob autoimmune, siv, piv txwv li, PCR-RFLPs, sequencing, los yog genotyping chips, ib rooj plaub ntawm RNA methylation tsis suav nrog. Nws yog ib qho tseem ceeb yog li ntawd kom paub lub xeev ntawm methylation ntawm interacting dsRNA thiab nws cov nyhuv ntawm allele MDA5. Raws li nyob rau hauv cov ntaub ntawv ntawm tus kab mob dsRNA kev lees paub (Wu li al. 2013) thiab qhov sib txawv ntawm tus kheej thiab tsis yog tus kheej mRNA (Züst et al. 2011), qhov poob ntawm ib pawg methyl los ntawm kev sib cuam tshuam MDA5, xws li hauv cov ntaub ntawv mutant Ile923Val MDA5, tej zaum yuav cuam tshuam cov filament tsim thiab induction ntawm hom I interferon.

Nco ntsoov, Plenge et al. (2013) yav dhau los tau tham txog lub peev xwm ntawm qhov tsis tshua muaj qhov sib txawv hauv cov kab mob ua rau cov noob los sawv cev rau lub hom phiaj kho mob rau kev cuam tshuam tshuaj. Txog rau qhov kawg no, kev ua haujlwm lom neeg ntawm cov kab mob sib txawv yuav tsum tau paub hauv txhua qhov kev sim los txuas cov kev tshawb nrhiav caj ces nrog lub hom phiaj kho tshiab. Yog li ntawd, nrhiav txoj hauj lwm ntawm qhov ua rau muaj kev sib txawv hauv 3D qauv ntawm cov protein thiab tshawb xyuas nws lub luag haujlwm los ntawm cov txheej txheem / kev ua haujlwm pom hauv txoj hauv kev pathogenetic ua rau tus kab mob autoimmune zoo li yog qhov tseem ceeb rau kev tswj xyuas ntxiv thiab kev kho mob zoo dua. cov neeg mob. Tseem muaj qhov tseem ceeb tshaj plaws uas yuav tsum tau txav mus dhau qhov kev tshawb pom ntawm SNPs cuam tshuam rau kev nkag siab tob dua ntawm cov kab mob sib txawv los piav qhia cov txheej txheem molecular thiab txoj hauv kev ntawm cov kab mob. Kev soj ntsuam kev ua haujlwm ntxiv yog xav tau los tshawb xyuas qhov kev sib txuas ntawm ribose-2′-Ο-methylated self-dsRNA rau MDA5 thiab qhov no cuam tshuam li cas MDA5 fibril sib dhos. Lub luag haujlwm tsis txaus ntseeg ntawm 2′-Ο-methylated ntawm mRNA raws li ib qho kev sib txawv ntawm tus kab mob thiab tus kheej-mRNA nyob rau hauv induction ntawm hom I interferon tej zaum yuav tau txuas mus rau ib tug tiv thaiv sensor nyob rau hauv tej yam autoimmunopathies.

Xaus

Qhov tsawg rs35667974 IFIH1 noob polymorphism tiv thaiv los ntawm T1D, PS, thiab PsA, thaum lub IFIH1 allele nqa los ntawm feem coob ntawm cov pejxeem predisposes rau cov kab mob. Cov nyiaj uas cov mutant Ile923Val MDA5 ua haujlwm sib txawv hauv kev sib cuam tshuam nrog tus kheej-dsRNA thiab tshwj xeeb nrog 2'-O-methylated ib qho qhia tias qee qhov sib txawv cuam tshuam nrog methylated dsRNA cuam tshuam MDA5 filament tsim, MAVS kev cuam tshuam, thiab filament tsim, thiab IFN signaling, raws li nyob rau hauv tus tswv tsev antiviral teb, tej zaum yuav tau raug xaiv tsis zoo vim hais tias lawv muaj kev tiv thaiv los ntawm cov kab mob.

Sau cov nyiaj pab

Conceptualization—AA, AP, EEE, thiab GNG, methodology—AA, AP, thiab EEE, validation—AA, AP, MIZ, GNG, thiab EEE, kev tshawb nrhiav—GNG, AA, AP, thiab MIZ, cov kev pab—GNG, AA, AP, thiab MIZ, cov ntaub ntawv curation—AA thiab AP, sau thiab thawj daim ntawv npaj-EEE, GNG, AA, AP, thiab MZ, sau, tshuaj xyuas, thiab kho cov ntawv sau—AA, AP, MZ, GNG, thiab EEE, Kev pom kev-AA, AP, thiab EEE, kev saib xyuas-EEE, thiab kev nrhiav nyiaj txiag-EEE. Txhua tus kws sau ntawv tau nyeem thiab pom zoo rau cov ntawv luam tawm ntawm cov ntawv sau.

Nyiaj txiag

Cov nyiaj qhib qhib yog muab los ntawm HEAL-Link Greece. Txoj haujlwm no tau txais kev txhawb nqa los ntawm qhov project "INSPIRED-The National Research Infrastructures on Integrated Structural Biology, Drug Screening Eforts and Drug Target Functional Characterization" (Grant MIS 5002550), uas yog ua raws li Txoj Cai "Reinforcement of the Research and Innovation Infrastructure," tau txais nyiaj los ntawm Txoj Haujlwm Kev Ua Haujlwm "Kev Sib Tw, Kev Ua Lag Luam, thiab Kev Tsim Kho Tshiab" (NSRF 2014-2020) thiab koom tes nyiaj txiag los ntawm Greece thiab European Union (European Regional Development Fund).

Cov ntaub ntawv muaj

Cov ntaub ntawv siv thiab/lossis tshuaj xyuas thaum lub sijhawm kawm tam sim no muaj los ntawm tus kws sau ntawv raws li kev thov.

Cov lus tshaj tawm

Kev tsis sib haum xeeb

Cov kws sau ntawv tshaj tawm tsis muaj kev sib cav txog kev txaus siab. Cov peev nyiaj tsis muaj lub luag haujlwm hauv kev tsim qauv ntawm kev kawm; hauv kev sau, txheeb xyuas, lossis txhais cov ntaub ntawv; hauv kev sau cov ntawv sau, lossis hauv kev txiav txim siab tshaj tawm cov txiaj ntsig.

Kev pom zoo rau kev ncaj ncees

Tsis siv tau.

Tso cai rau koom

Tsis siv tau.

Kev tso cai rau kev tshaj tawm

Tsis siv tau.

Qhib Access

Kab lus no tau tso cai raws li Creative Commons Attribution 4.0 Daim ntawv tso cai thoob ntiaj teb, uas tso cai siv, sib koom, hloov kho, faib, thiab luam tawm hauv ib qho nruab nrab lossis hom ntawv, tsuav yog koj muab credit rau tus thawj sau ) thiab qhov chaw, muab qhov txuas mus rau Creative Commons daim ntawv tso cai, thiab qhia seb puas tau hloov pauv. Cov duab lossis lwm yam khoom siv thib peb hauv tsab xov xwm no suav nrog hauv tsab xov xwm Creative Commons daim ntawv tso cai tshwj tsis yog tau qhia lwm yam hauv kab qiv nyiaj rau cov khoom siv. Piv txwv tias cov ntaub ntawv tsis suav nrog hauv tsab xov xwm Creative Commons daim ntawv tso cai thiab koj qhov kev npaj siv tsis raug tso cai los ntawm txoj cai lij choj lossis tshaj qhov kev tso cai siv. Nyob rau hauv tas li ntawd, koj yuav tsum tau txais kev tso cai ncaj qha los ntawm lub copyright tuav.

Cov ntaub ntawv

1. Aduri R, Psciuk BT, Saro P et al (2007) AMBER force field parameters for the naturally happening modified nucleosides in RNA. J Chem Theory Comput 3:1464–1475.

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