Identification of novel Arabidopsis thaliana upstream open reading frames that control expression of the main coding sequences in a peptide sequence-dependent manner.
Identification of novel Arabidopsis thaliana upstream open reading frames that control expression of the main coding sequences in a peptide sequence-dependent manner.
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DOI：
10.1093/nar/gkv018
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发表时间：
2015-02-18
影响因子：
14.9
通讯作者：
Onouchi H
中科院分区：
文献类型：
--
作者：
Ebina I;Takemoto-Tsutsumi M;Watanabe S;Koyama H;Endo Y;Kimata K;Igarashi T;Murakami K;Kudo R;Ohsumi A;Noh AL;Takahashi H;Naito S;Onouchi H
上游开放阅读框（uORF）通常存在于真核生物mRNA的5′-前导区，可负调控下游主ORF的翻译效率。尽管大多数uORF的作用被认为与其编码的肽序列无关，但某些uORF以肽序列依赖性方式控制主要ORF的翻译。为了全基因组鉴定这种肽序列依赖性调控uORF，已经使用生物信息学分析对具有保守氨基酸序列的uORF进行了详尽的搜索。然而，这些生物信息学方法鉴定的保守的uORF是否编码调节肽还没有实验确定。在这里，我们分析了16个最近确定的拟南芥保守的uORF的影响，其氨基酸序列的主要ORF的表达，使用瞬时表达试验。我们确定了五个新的uORF，抑制主要ORF表达的肽序列依赖性的方式。突变分析表明，在其中四个，C-末端区域的uORF编码的肽是关键的主要ORF表达的抑制。有趣的是，我们还确定了一个特殊的序列依赖性调控uORF，其中终止密码子的位置是不保守的，C-末端区域是不重要的主要ORF表达的抑制。
Upstream open reading frames (uORFs) are often found in the 5′-leader regions of eukaryotic mRNAs and can negatively modulate the translational efficiency of the downstream main ORF. Although the effects of most uORFs are thought to be independent of their encoded peptide sequences, certain uORFs control translation of the main ORF in a peptide sequence-dependent manner. For genome-wide identification of such peptide sequence-dependent regulatory uORFs, exhaustive searches for uORFs with conserved amino acid sequences have been conducted using bioinformatic analyses. However, whether the conserved uORFs identified by these bioinformatic approaches encode regulatory peptides has not been experimentally determined. Here we analyzed 16 recently identified Arabidopsis thaliana conserved uORFs for the effects of their amino acid sequences on the expression of the main ORF using a transient expression assay. We identified five novel uORFs that repress main ORF expression in a peptide sequence-dependent manner. Mutational analysis revealed that, in four of them, the C-terminal region of the uORF-encoded peptide is critical for the repression of main ORF expression. Intriguingly, we also identified one exceptional sequence-dependent regulatory uORF, in which the stop codon position is not conserved and the C-terminal region is not important for the repression of main ORF expression.
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影响因子：
16
作者：
Chiba, Shinobu;Ito, Koreaki
通讯作者：
Ito, Koreaki
影响因子：
4.8
作者：
Hanfrey, C;Elliott, KA;Michael, AJ
通讯作者：
Michael, AJ
影响因子：
5.4
作者：
Hayden CA;Jorgensen RA
通讯作者：
Jorgensen RA
影响因子：
16
作者：
Cruz-Vera, LR;Rajagopal, S;Yanofsky, C
通讯作者：
Yanofsky, C
影响因子：
7.8
作者：
BLOBEL, G;SABATINI, DD
通讯作者：
SABATINI, DD