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大肠杆菌中改变的mRNA稳定性(ams)基因分析。核苷酸序列、转录分析及其产物与粗糙脉孢菌线粒体核糖体蛋白MRP3的同源性。

Analysis of the altered mRNA stability (ams) gene from Escherichia coli. Nucleotide sequence, transcriptional analysis, and homology of its product to MRP3, a mitochondrial ribosomal protein from Neurospora crassa.

作者信息

Claverie-Martin F, Diaz-Torres M R, Yancey S D, Kushner S R

机构信息

Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

出版信息

J Biol Chem. 1991 Feb 15;266(5):2843-51.

PMID:1704367
Abstract

The product of the altered mRNA stability (ams) gene of Escherichia coli is involved in decay of mRNA. The complete nucleotide sequence of a 4-kilobase BamHI restriction fragment containing the ams coding sequence was determined. Transcription of the ams gene was analyzed by high resolution S1 mapping. A promoter was found with a homology score of 58% 361 nucleotides upstream from the start codon of ams. The ams structural gene consists of an open reading frame of 2,445 nucleotides. The protein predicted from this open reading frame has a molecular mass of 91,327 Da, which is significantly smaller than that determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Confirmation of the ams coding sequence was obtained by comparing the predicted amino acid sequence with that derived by amino-terminal analysis of gel-purified Ams protein. The predicted protein sequence of the ams gene was screened against translations of the GenBank DNA sequence data base. A homology of 18% over a region of 315 amino acids of the carboxyl terminus of the Ams product was found to MRP3, a mitochondrial ribosomal protein from Neurospora crassa. A smaller region of homology (29% in 86 residues) was found to the human U1 small nuclear ribonucleoparticle 70,000-Da protein.

摘要

大肠杆菌中改变的mRNA稳定性(ams)基因的产物参与mRNA的降解。测定了包含ams编码序列的4千碱基BamHI限制性片段的完整核苷酸序列。通过高分辨率S1作图分析ams基因的转录。在ams起始密码子上游361个核苷酸处发现了一个同源性得分58%的启动子。ams结构基因由一个2445个核苷酸的开放阅读框组成。从这个开放阅读框预测的蛋白质分子量为91327道尔顿,明显小于通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析确定的分子量。通过将预测的氨基酸序列与凝胶纯化的Ams蛋白的氨基末端分析得到的序列进行比较,证实了ams编码序列。针对GenBank DNA序列数据库的翻译筛选ams基因的预测蛋白质序列。发现Ams产物羧基末端315个氨基酸区域与粗糙脉孢菌的线粒体核糖体蛋白MRP3有18%的同源性。发现与人类U1小核核糖核蛋白70000道尔顿蛋白有较小的同源区域(86个残基中有29%)。

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