Macario A J, Simon V H, Conway de Macario E
Wadsworth Center, New York State Department of Health, Albany 12201-0509, USA.
Biochim Biophys Acta. 1995 Nov 7;1264(2):173-7. doi: 10.1016/0167-4781(95)00163-b.
In some eubacteria with a dnaK locus in which grpE is close upstream of dnaK, grpE is preceded by an open reading frame (orf) believed to be a heat-shock gene. We also found an orf, orf16, upstream of grpE in the archaeon Methanosarcina mazei S-6, but this gene differs from the eubacterial counterpart: it is shorter, does not respond to a temperature upshift as heat-shock genes do, and the deduced protein Orf16, does not resemble the proteins coded by the eubacterial equivalents. orf16 is expressed monocistronically, with a transcription initiation site 24 bases upstream of the translation start codon, 22 bases downstream of a putative promoter identical to the consensus promoter for genes in methanogens. This initiation site is used by heat-shocked and non-heat-shocked cells in the two morphologic stages of M. mazei S-6 tested, i.e., packets and single cells. Three transcription termination sites were identified, one of which is detectable only in non-heat-shocked cells. Data from comparative analyses of the Orf16 deduced amino acid sequence and those of other known proteins, as well as the apparent biochemical characteristics of Orf16, suggest that the latter is a membrane molecule.
在一些具有dnaK基因座的真细菌中,grpE位于dnaK的紧邻上游,grpE之前有一个开放阅读框(orf),被认为是一个热休克基因。我们还在古菌马氏甲烷八叠球菌S-6的grpE上游发现了一个orf,即orf16,但该基因与真细菌中的对应基因不同:它较短,不像热休克基因那样对温度升高作出反应,并且推导的蛋白质Orf16与真细菌中对应基因编码的蛋白质不相似。orf16以单顺反子形式表达,转录起始位点在翻译起始密码子上游24个碱基处,在一个与产甲烷菌基因的共有启动子相同的假定启动子下游22个碱基处。在所测试的马氏甲烷八叠球菌S-6的两个形态阶段,即细胞团和单细胞中,热休克和非热休克细胞均使用该起始位点。鉴定出了三个转录终止位点,其中一个仅在非热休克细胞中可检测到。对Orf16推导氨基酸序列与其他已知蛋白质的氨基酸序列进行比较分析的数据,以及Orf16明显的生化特性表明,后者是一种膜分子。