Klumpp D J, Plank D W, Bowdin L J, Stueland C S, Chung T, LaPorte D C
Department of Biochemistry, University of Minnesota, Minneapolis 55455.
J Bacteriol. 1988 Jun;170(6):2763-9. doi: 10.1128/jb.170.6.2763-2769.1988.
In Escherichia coli, the phosphorylation and dephosphorylation of isocitrate dehydrogenase (IDH) are catalyzed by a bifunctional protein kinase/phosphatase. We have determined the nucleotide sequence of aceK, the gene encoding IDH kinase/phosphatase. This gene consists of a single open reading frame of 1,734 base pairs preceded by a Shine-Dalgarno ribosome-binding site. Examination of the deduced amino acid sequence of IDH kinase/phosphatase revealed sequences which are similar to the consensus sequence for ATP-binding sites. This protein did not, however, exhibit the extensive sequence homologies which are typical of other protein kinases. Multiple copies of the REP family of repetitive extragenic elements were found within the intergenic region between aceA (encoding isocitrate lyase) and aceK. These elements have the potential for combining to form an exceptionally stable stem-loop structure (delta G = -54 kcal/mol [ca. -226 kJ/mol]) in the mRNA. This structure, which masks the ribosome-binding site and start codon for aceK, may contribute to the downshift in expression observed between aceA and aceK. Another potential stem-loop structure (delta G = -29 kcal/mol [ca. 121 kJ/mol]), unrelated to the REP sequences, was found within aceK.
在大肠杆菌中,异柠檬酸脱氢酶(IDH)的磷酸化和去磷酸化由一种双功能蛋白激酶/磷酸酶催化。我们已经确定了编码IDH激酶/磷酸酶的基因aceK的核苷酸序列。该基因由一个1734个碱基对的单一开放阅读框组成,前面有一个Shine-Dalgarno核糖体结合位点。对推导的IDH激酶/磷酸酶氨基酸序列的检查揭示了与ATP结合位点的共有序列相似的序列。然而,这种蛋白质并没有表现出其他蛋白激酶典型的广泛序列同源性。在编码异柠檬酸裂合酶的aceA和aceK之间的基因间隔区内发现了多个重复基因外元件REP家族的拷贝。这些元件有可能在mRNA中结合形成一个异常稳定的茎环结构(ΔG = -54千卡/摩尔[约-226千焦/摩尔])。这种结构掩盖了aceK的核糖体结合位点和起始密码子,可能导致在aceA和aceK之间观察到的表达下调。在aceK内发现了另一种与REP序列无关的潜在茎环结构(ΔG = -29千卡/摩尔[约121千焦/摩尔])。