Crowell D N, Reznikoff W S, Raetz C R
Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison 53706.
J Bacteriol. 1987 Dec;169(12):5727-34. doi: 10.1128/jb.169.12.5727-5734.1987.
The lpxB gene of Escherichia coli, believed to be the structural gene for lipid A disaccharide synthase, is located in the min 4 region of the chromosome. It is adjacent to and clockwise of the lpxA gene, which is thought to encode UDP-N-acetylglucosamine acyltransferase. Preliminary evidence suggests that lpxA and lpxB are cotranscribed in the clockwise direction and thus constitute part of a previously unknown operon (D. N. Crowell, M. S. Anderson, and C. R. H. Raetz, J. Bacteriol. 168:152-159, 1986). We now report the complete nucleotide sequence of a 1,522-base-pair PvuII-HincII fragment known to carry the lpxB gene. This sequence contained an open reading frame of 1,149 base pairs, in agreement with the predicted size, location, and orientation of lpxB. There was a second open reading frame 5' to, and in the same orientation as, lpxB that corresponded to lpxA. The ochre codon terminating lpxA was shown to overlap the methionine codon identified as the initiation codon for lpxB, suggesting that these genes are cotranscribed and translationally coupled. A third open reading frame was also shown to begin at the 3' end of lpxB with analogous overlap between the opal codon terminating lpxB and the methionine codon that putatively initiates translation downstream of lpxB in the clockwise direction. These results argue that at least three genes constitute a translationally coupled operon in the min 4 region of the E. coli chromosome. The accompanying paper by Tomasiewicz and McHenry (J. Bacteriol. 169:5735-5744, 1987) presents 4.35 kilobases of DNA sequence, beginning at the 3' end of lpxB, and argues that dnaE and several other open reading frames may be members of this operon.
大肠杆菌的lpxB基因被认为是脂多糖A二糖合酶的结构基因,位于染色体的4分钟区域。它与lpxA基因相邻且在其顺时针方向,lpxA基因被认为编码UDP-N-乙酰葡糖胺酰基转移酶。初步证据表明,lpxA和lpxB以顺时针方向共转录,因此构成了一个先前未知的操纵子的一部分(D.N.克罗威尔、M.S.安德森和C.R.H.雷茨,《细菌学杂志》168:152 - 159,1986年)。我们现在报告一个已知携带lpxB基因的1522碱基对的PvuII - HincII片段的完整核苷酸序列。该序列包含一个1149碱基对的开放阅读框,与lpxB预测的大小、位置和方向一致。在lpxB的5'端且与lpxB方向相同有第二个开放阅读框,它对应于lpxA。终止lpxA的赭石密码子显示与被确定为lpxB起始密码子的甲硫氨酸密码子重叠,这表明这些基因是共转录且翻译偶联的。还显示第三个开放阅读框从lpxB的3'端开始,终止lpxB的乳白密码子与假定在顺时针方向lpxB下游起始翻译的甲硫氨酸密码子之间有类似的重叠。这些结果表明,至少三个基因在大肠杆菌染色体的4分钟区域构成一个翻译偶联的操纵子。托马谢维茨和麦克亨利的随附论文(《细菌学杂志》169:5735 - 5744,1987年)展示了从lpxB的3'端开始的4.35千碱基的DNA序列,并认为dnaE和其他几个开放阅读框可能是这个操纵子的成员。