Inosine modifications in human tRNAs are incorporated at the precursor tRNA level.
Inosine modifications in human tRNAs are incorporated at the precursor tRNA level.
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DOI：
10.1093/nar/gkv277
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发表时间：
2015-05-26
影响因子：
14.9
通讯作者：
Ribas de Pouplana L
中科院分区：
文献类型：
--
作者：
Torres AG;Piñeyro D;Rodríguez-Escribà M;Camacho N;Reina O;Saint-Léger A;Filonava L;Batlle E;Ribas de Pouplana L
转移rna （trna）是遗传密码的关键适配分子，在转录后被大量修饰。反密码子第一个残基上的肌苷（位置34；I34）是一种重要的广泛存在的tRNA修饰，迄今为止研究较少。真核生物的这种修饰是由异二聚体酶腺苷脱氨酶作用于tRNA （hetADAT）催化的腺嘌呤脱氨反应引起的，该反应由两个亚基组成：ADAT2和ADAT3。利用高通量小RNA测序（RNAseq），我们发现这种修饰在前体tRNA水平和成熟过程中被纳入人类tRNA。我们还从功能上验证了编码heatat的人类基因，并表明该酶的亚基以adat2依赖的方式在细胞核中共定位。最后，通过敲除HsADAT2，我们证明了hetADAT细胞水平的变化将导致其所有潜在底物中I34修饰水平的变化。总之，我们将RNAseq作为一个强大的工具，在前体tRNA水平上研究转录后tRNA修饰，并首次深入了解后生动物中I34 tRNA修饰的生物学原理。
Transfer RNAs (tRNAs) are key adaptor molecules of the genetic code that are heavily modified post-transcriptionally. Inosine at the first residue of the anticodon (position 34; I34) is an essential widespread tRNA modification that has been poorly studied thus far. The modification in eukaryotes results from a deamination reaction of adenine that is catalyzed by the heterodimeric enzyme adenosine deaminase acting on tRNA (hetADAT), composed of two subunits: ADAT2 and ADAT3. Using high-throughput small RNA sequencing (RNAseq), we show that this modification is incorporated to human tRNAs at the precursor tRNA level and during maturation. We also functionally validated the human genes encoding for hetADAT and show that the subunits of this enzyme co-localize in nucleus in an ADAT2-dependent manner. Finally, by knocking down HsADAT2, we demonstrate that variations in the cellular levels of hetADAT will result in changes in the levels of I34 modification in all its potential substrates. Altogether, we present RNAseq as a powerful tool to study post-transcriptional tRNA modifications at the precursor tRNA level and give the first insights on the biology of I34 tRNA modification in metazoans.
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影响因子：
14.9
作者：
Ebhardt HA;Tsang HH;Dai DC;Liu Y;Bostan B;Fahlman RP
通讯作者：
Fahlman RP
影响因子：
56.9
作者：
Gerber, AP;Keller, W
通讯作者：
Keller, W
影响因子：
14.9
作者：
Flicek P;Amode MR;Barrell D;Beal K;Billis K;Brent S;Carvalho-Silva D;Clapham P;Coates G;Fitzgerald S;Gil L;Girón CG;Gordon L;Hourlier T;Hunt S;Johnson N;Juettemann T;Kähäri AK;Keenan S;Kulesha E;Martin FJ;Maurel T;McLaren WM;Murphy DN;Nag R;Overduin B;Pignatelli M;Pritchard B;Pritchard E;Riat HS;Ruffier M;Sheppard D;Taylor K;Thormann A;Trevanion SJ;Vullo A;Wilder SP;Wilson M;Zadissa A;Aken BL;Birney E;Cunningham F;Harrow J;Herrero J;Hubbard TJ;Kinsella R;Muffato M;Parker A;Spudich G;Yates A;Zerbino DR;Searle SM
通讯作者：
Searle SM
影响因子：
7
作者：
Harrow J;Frankish A;Gonzalez JM;Tapanari E;Diekhans M;Kokocinski F;Aken BL;Barrell D;Zadissa A;Searle S;Barnes I;Bignell A;Boychenko V;Hunt T;Kay M;Mukherjee G;Rajan J;Despacio-Reyes G;Saunders G;Steward C;Harte R;Lin M;Howald C;Tanzer A;Derrien T;Chrast J;Walters N;Balasubramanian S;Pei B;Tress M;Rodriguez JM;Ezkurdia I;van Baren J;Brent M;Haussler D;Kellis M;Valencia A;Reymond A;Gerstein M;Guigó R;Hubbard TJ
通讯作者：
Hubbard TJ
影响因子：
14.9
作者：
GROSJEAN, H;CONSTANTINESCO, F;BENACHENHOU, N
通讯作者：
BENACHENHOU, N