Keeling Kim M, Salas-Marco Joe, Osherovich Lev Z, Bedwell David M
Department of Microbiology, BBRB 432/Box 8, 1530 3rd Avenue South, University of Alabama at Birmingham, Birmingham, AL 35294-2170, USA.
Mol Cell Biol. 2006 Jul;26(14):5237-48. doi: 10.1128/MCB.02448-05.
In this report, we show that the Saccharomyces cerevisiae protein Tpa1p (for termination and polyadenylation) influences translation termination efficiency, mRNA poly(A) tail length, and mRNA stability. Tpa1p is encoded by the previously uncharacterized open reading frame YER049W. Yeast strains carrying a deletion of the TPA1 gene (tpa1Delta) exhibited increased readthrough of stop codons, and coimmunoprecipitation assays revealed that Tpa1p interacts with the translation termination factors eRF1 and eRF3. In addition, the tpa1Delta mutation led to a 1.5- to 2-fold increase in the half-lives of mRNAs degraded by the general 5'-->3' pathway or the 3'-->5' nonstop decay pathway. In contrast, this mutation did not have any affect on the nonsense-mediated mRNA decay pathway. Examination of mRNA poly(A) tail length revealed that poly(A) tails are longer than normal in a tpa1Delta strain. Consistent with a potential role in regulating poly(A) tail length, Tpa1p was also found to coimmunoprecipitate with the yeast poly(A) binding protein Pab1p. These results suggest that Tpa1p is a component of a messenger ribonucleoprotein complex bound to the 3' untranslated region of mRNAs that affects translation termination, deadenylation, and mRNA decay.
在本报告中,我们表明酿酒酵母蛋白Tpa1p(用于终止和聚腺苷酸化)影响翻译终止效率、mRNA聚(A)尾长度和mRNA稳定性。Tpa1p由先前未鉴定的开放阅读框YER049W编码。携带TPA1基因缺失(tpa1Δ)的酵母菌株表现出终止密码子通读增加,共免疫沉淀试验表明Tpa1p与翻译终止因子eRF1和eRF3相互作用。此外,tpa1Δ突变导致通过一般的5'→3'途径或3'→5'无义衰变途径降解的mRNA半衰期增加1.5至2倍。相比之下,该突变对无义介导的mRNA衰变途径没有任何影响。对mRNA聚(A)尾长度的检查表明,在tpa1Δ菌株中聚(A)尾比正常情况更长。与调节聚(A)尾长度的潜在作用一致,还发现Tpa1p与酵母聚(A)结合蛋白Pab1p共免疫沉淀。这些结果表明,Tpa1p是信使核糖核蛋白复合物的一个组成部分,该复合物与mRNA的3'非翻译区结合,影响翻译终止、去腺苷酸化和mRNA衰变。