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首页> 美国卫生研究院文献>Genetics >Allele-Specific Suppressors of lin-1(R175Opal) Identify Functions of MOC-3 and DPH-3 in tRNA Modification Complexes in Caenorhabditis elegans
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Allele-Specific Suppressors of lin-1(R175Opal) Identify Functions of MOC-3 and DPH-3 in tRNA Modification Complexes in Caenorhabditis elegans

机译：lin-1（R175Opal）的等位基因特异性抑制剂可鉴定秀丽隐杆线虫tRNA修饰复合物中MOC-3和DPH-3的功能。

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摘要

The elongator (ELP) complex consisting of Elp1-6p has been indicated to play roles in multiple cellular processes. In yeast, the ELP complex has been shown to genetically interact with Uba4p/Urm1p and Kti11-13p for a function in tRNA modification. Through a Caenorhabditis elegans genetic suppressor screen and positional cloning, we discovered that loss-of-function mutations of and , orthologs of the yeast UBA4 and KTI11, respectively, effectively suppress the Multivulva () phenotype of the (, R175Opal) mutation. These mutations do not suppress the phenotype caused by other alleles or by gain-of-function alleles of ras or raf that act upstream of . The suppression can also be reverted by RNA interference of . Furthermore, we showed that (lf) also suppressed the defect of () in promoting the expression of a downstream target (). These results indicate that suppression by the and mutations is due to the elevated activity of (href="http://www.wormbase.org/db/get?name=e1275;class=Variation" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">e1275) itself rather than the altered activity of a factor downstream of href="http://www.wormbase.org/db/get?name=lin-1;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">lin-1. We further showed that loss-of-function mutations of href="http://www.wormbase.org/db/get?name=urm-1;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">urm-1 and href="http://www.wormbase.org/db/get?name=elpc-1;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">elpc-1href="http://www.wormbase.org/db/get?name=elpc-4;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">-4, the worm counterparts of URM1 and ELP complex components in yeast, also suppressed href="http://www.wormbase.org/db/get?name=lin-1;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">lin-1(href="http://www.wormbase.org/db/get?name=e1275;class=Variation" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">e1275). We also confirmed that href="http://www.wormbase.org/db/get?name=moc-3;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">moc-3(lf) and href="http://www.wormbase.org/db/get?name=dph-3;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">dph-3(lf) have defects in tRNA modifications as do the mutants of their yeast orthologs. These results, together with the observation of a likely readthrough product from a lin-1(e1275)∷gfp fusion transgene indicate that the aberrant tRNA modification led to failed recognition of a premature stop codon in href="http://www.wormbase.org/db/get?name=lin-1;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">lin-1(href="http://www.wormbase.org/db/get?name=e1275;class=Variation" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">e1275). Our genetic data suggest that the functional interaction of href="http://www.wormbase.org/db/get?name=moc-3;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">moc-3/href="http://www.wormbase.org/db/get?name=urm-1;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">urm-1 and href="http://www.wormbase.org/db/get?name=dph-3;class=Gene" data-ga-action="click_feat_suppl" ref="reftype=extlink&article-id=2927752&issue-id=189230&journal-id=301&FROM=Article%7CFront%20Matter&TO=External%7CLink%7CURI" target="_blank">dph-3 with the ELP complex is an evolutionarily conserved mechanism involved in tRNA functions that are important for accurate translation.
机译：已经表明由Elp1-6p组成的延伸子（ELP）复合物在多个细胞过程中发挥作用。在酵母中，ELP复合物已显示与Uba4p / Urm1p和Kti11-13p发生遗传相互作用，从而具有tRNA修饰的功能。通过秀丽隐杆线虫遗传抑制基因筛选和位置克隆，我们发现酵母UBA4和KTI11的和的直系同源物的功能缺失突变分别有效地抑制了（R175Opal）突变的Multivulva（）表型。这些突变不会抑制由其他等位基因或ras或raf上游起作用的ras或raf的功能获得等位基因引起的表型。抑制也可以被RNAi干扰恢复。此外，我们显示（lf）还抑制（）促进下游靶标（）表达的缺陷。这些结果表明被 和 突变抑制的原因是（href =“ http://www.wormbase.org/db / get？name = e1275; class = Variation“ data-ga-action =” click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI “ target =” _ blank“> e1275 ）本身，而不是 href =” http://www.wormbase.org/db/get？ name = lin-1; class = Gene“ data-ga-action =” click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =“ _ blank”> lin-1 。我们进一步显示 href =“ http://www.wormbase.org/db/get?name=urm-1;class=Gene” data-ga-action =“的功能丧失突变click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> urm-1 和 href =“ http://www.wormbase.org/db/get?name=elpc-1;class=Gene” data-ga-action =“ click_feat_suppl” ref =“ reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> elpc-1 href =” http://www.wormbase.org / db / get？name = elpc-4; class = Gene“ data-ga-action =” click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External ％7CLink％7CURI“ target =” _ blank“>-4 （酵母中 URM1 和ELP复杂成分的蠕虫对应物）也抑制了 href = “ http://www.wormbase.org/db/get?name=lin-1;class=Gene” data-ga-action =“ click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> lin-1 （href =“ http://www.wormbase.org/db/get?name=e1275;class=Variation” data-ga-action =“ click_feat_suppl” ref =“ reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> e1275 ）。我们还确认 href =“ http://www.wormbase.org/db/get?name=moc-3;class=Gene” data-ga-action =“ click_feat_suppl” ref =“ reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> moc-3 （lf）和 href =“ http://www.wormbase.org/db/get?name=dph-3;class=Gene” data-ga-action =“ click_feat_suppl” ref =“ reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> dph-3 （lf）的tRNA修饰存在缺陷，其突变体也是如此酵母直系同源物。这些结果，并观察到 lin-1（e1275）∷gfp融合转基因的可能通读产物，表明异常的tRNA修饰导致 lin-1 （href =” http://www.wormbase.org/db/get？ name = e1275; class = Variation“ data-ga-action =” click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target = “ _blank”> e1275 ）。我们的遗传数据表明 href =“ http://www.wormbase.org/db/get?name=moc-3;class=Gene” data-ga-action =“ click_feat_suppl”的功能相互作用ref =“ reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI” target =“ _ blank”> moc-3 / < em> href =“ http://www.wormbase.org/db/get?name=urm-1;class=Gene” data-ga-action =“ click_feat_suppl” ref =“ reftype = extlink＆article-id = 2927752＆issue -id = 189230＆journal-id = 301＆FROM = Article％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> urm-1 和 href =” http：/ /www.wormbase.org/db/get?name=dph-3;class=Gene“ data-ga-action =” click_feat_suppl“ ref =” reftype = extlink＆article-id = 2927752＆issue-id = 189230＆journal-id = 301＆FROM = Article与ELP配合使用的％7CFront％20Matter＆TO = External％7CLink％7CURI“ target =” _ blank“> dph-3 是tRNA功能涉及的进化保守机制，对于准确翻译非常重要。

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