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一个大肠杆菌操纵子的核苷酸序列,该操纵子包含tRNA(m1G)甲基转移酶、核糖体蛋白S16和L19以及一种21K多肽的基因。

The nucleotide sequence of an Escherichia coli operon containing genes for the tRNA(m1G)methyltransferase, the ribosomal proteins S16 and L19 and a 21-K polypeptide.

作者信息

Byström A S, Hjalmarsson K J, Wikström P M, Björk G R

出版信息

EMBO J. 1983;2(6):899-905. doi: 10.1002/j.1460-2075.1983.tb01519.x.

Abstract

The nucleotide sequence of a 4.6-kb SalI-EcoRI DNA fragment including the trmD operon, located at min 56 on the Escherichia coli K-12 chromosome, has been determined. The trmD operon encodes four polypeptides: ribosomal protein S16 (rpsP), 21-K polypeptide (unknown function), tRNA-(m1G)methyltransferase (trmD) and ribosomal protein L19 (rplS), in that order. In addition, the 4.6-kb DNA fragment encodes a 48-K and a 16-K polypeptide of unknown functions which are not part of the trmD operon. The mol. wt. of tRNA(m1G)methyltransferase determined from the DNA sequence is 28 424. The probable locations of promoter and terminator of the trmD operon are suggested. The translational start of the trmD gene was deduced from the known NH2-terminal amino acid sequence of the purified enzyme. The intercistronic regions in the operon vary from 9 to 40 nucleotides, supporting the earlier conclusion that the four genes are co-transcribed, starting at the major promoter in front of the rpsP gene. Since it is known that ribosomal proteins are present at 8000 molecules/genome and the tRNA-(m1G)methyltransferase at only approximately 80 molecules/genome in a glucose minimal culture, some powerful regulatory device must exist in this operon to maintain this non-coordinate expression. The codon usage of the two ribosomal protein genes is similar to that of other ribosomal protein genes, i.e., high preference for the most abundant tRNA isoaccepting species. The trmD gene has a codon usage typical for a protein made in low amount in accordance with the low number of tRNA-(m1G)methyltransferase molecules found in the cell.

摘要

已确定位于大肠杆菌K-12染色体56分钟处、包含trmD操纵子的一个4.6kb SalI-EcoRI DNA片段的核苷酸序列。trmD操纵子依次编码四种多肽:核糖体蛋白S16(rpsP)、21K多肽(功能未知)、tRNA-(m1G)甲基转移酶(trmD)和核糖体蛋白L19(rplS)。此外,该4.6kb DNA片段还编码两种功能未知的48K和16K多肽,它们不属于trmD操纵子。根据DNA序列确定的tRNA(m1G)甲基转移酶的分子量为28424。文中还推测了trmD操纵子启动子和终止子的可能位置。trmD基因的翻译起始位点是根据纯化酶已知的NH2末端氨基酸序列推导出来的。操纵子中的基因间区从9到40个核苷酸不等,这支持了之前的结论,即这四个基因是从rpsP基因前面的主要启动子开始共转录的。已知在葡萄糖基本培养基中,核糖体蛋白在每个基因组中有8000个分子,而tRNA-(m1G)甲基转移酶每个基因组中只有大约80个分子,因此该操纵子中必定存在某种强大的调控机制来维持这种非协调表达。两个核糖体蛋白基因的密码子使用情况与其他核糖体蛋白基因相似,即对最丰富的同功tRNA种类有高度偏好。trmD基因的密码子使用情况具有典型性,即根据细胞中发现的tRNA-(m1G)甲基转移酶分子数量较少,该基因编码的蛋白产量较低。

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