Hosted T J, Rochefort D A, Benson D R
Department of Molecular and Cell Biology, University of Connecticut, Storrs 06269-3044.
J Bacteriol. 1993 Jun;175(11):3679-84. doi: 10.1128/jb.175.11.3679-3684.1993.
Frankia alni CpI1 has two glutamine synthetases (GSs), GSI and GSII. The GSI gene (glnA) was isolated from a cosmid library of F. alni CpI1 DNA by heterologous probing with glnA from Streptomyces coelicolor. The glnA gene was shown to be located upstream of the GSII gene (glnII) by DNA-DNA hybridization. The nucleotide sequences of the 1,422-bp CpI1 glnA gene and of the 449-bp intervening region between glnA and glnII were determined, and the glnA amino acid sequence was deduced. In common with GSIs from other organisms, CpI1 GSI contains five conserved regions near the active site and a conserved tyrosine at the adenylylation site. F. alni CpI1 glnA complemented the glutamine growth requirement of the Escherichia coli glnA deletion strain YMC11 but only when expressed from an E. coli lac promoter. While the functional significance of maintaining two GSs adjacent to one another remains unclear, this arrangement in F. alni provides support for the recently proposed origin of GSI and GSII as resulting from a gene duplication early in the evolution of life.
桤木弗兰克氏菌CpI1有两种谷氨酰胺合成酶(GSs),即GSI和GSII。通过用天蓝色链霉菌的glnA进行异源探针杂交,从桤木弗兰克氏菌CpI1 DNA的黏粒文库中分离出GSI基因(glnA)。通过DNA-DNA杂交表明,glnA基因位于GSII基因(glnII)的上游。测定了1422 bp的CpI1 glnA基因以及glnA和glnII之间449 bp间隔区的核苷酸序列,并推导了glnA的氨基酸序列。与其他生物的GSI一样,CpI1 GSI在活性位点附近含有五个保守区域,在腺苷酸化位点含有一个保守的酪氨酸。桤木弗兰克氏菌CpI1 glnA补充了大肠杆菌glnA缺失菌株YMC11对谷氨酰胺的生长需求,但只有当从大肠杆菌lac启动子表达时才行。虽然维持两个相邻的GSs的功能意义尚不清楚,但桤木弗兰克氏菌中的这种排列为最近提出的GSI和GSII起源于生命进化早期的基因复制提供了支持。