Biochemical examination of the potential eukaryotic-type protein kinase genes in the complete genome of the unicellular Cyanobacterium synechocystis sp. PCC 6803.

A. Kamei, T. Yuasa, Xiaoxing Geng, M. Ikeuchi
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引用次数: 42

Abstract

The complete genome of the unicellular motile cyanobacterium Synechocystis sp. PCC 6803 harbors seven putative genes for a subfamily Pkn2 of the eukaryotic-type (or "Hanks-type") protein kinase. Previously, SpkA and SpkB were shown to have protein kinase activity and to be required for cell motility. Here, the other five genes were examined. These genes, except for spkG (slr0152), were successfully expressed in Escherichia coli. Eukaryotic-type protein kinase activity of the expressed SpkC (Slr0599), SpkD (S110776) and SpkF (Slr1225) was demonstrated as autophosphorylation and phosphorylation of the general substrate proteins. SpkE (Slr1443) did not show any activity, a finding consistent with its lack of several key amino acid residues in its kinase motif. Gene-disrupted mutants showed no discernible defect in phenotype except that spkD was apparently essential for survival.
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单细胞藻胞杆菌PCC 6803全基因组真核型蛋白激酶基因的生化检测。
单细胞活动蓝藻聚胞杆菌(synnechocystis sp. PCC 6803)的完整基因组包含真核型(或“汉克斯型”)蛋白激酶Pkn2亚家族的七个假定基因。以前,SpkA和SpkB被证明具有蛋白激酶活性,并且是细胞运动所必需的。在这里,其他五个基因被检查。除spkG (slr0152)外,其余基因均在大肠杆菌中成功表达。表达的SpkC (Slr0599), SpkD (S110776)和SpkF (Slr1225)的真核型蛋白激酶活性被证明是自磷酸化和磷酸化一般底物蛋白。SpkE (Slr1443)没有表现出任何活性,这与其激酶基序中缺乏几个关键氨基酸残基一致。基因破坏突变体在表型上没有明显的缺陷,除了spkD显然是生存所必需的。
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