Sprušanský O, Řežuchová B, Homerová D, Kormanec J
Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 842 51 Bratislava, Slovak Republic1.
Microbiology (Reading). 2001 May;147(Pt 5):1291-1301. doi: 10.1099/00221287-147-5-1291.
Expression of the gap gene encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is developmentally regulated, and induced by glucose in Streptomyces aureofaciens. A gene, gapR, encoding a protein similar to the AraC/XylS family of bacterial transcriptional regulators was identified upstream of gap. The gapR gene was constitutively expressed from a single promoter during the course of differentiation. By integrative transformation, via double crossover, a stable null mutant of the gapR gene was obtained. The mutation only slightly affected growth, and had no effect on differentiation of S. aureofaciens. However, transcription of the GAPDH-encoding gap gene was substantially reduced in the S. aureofaciens DeltagapR null mutant, irrespective of carbon source used. Though GAPDH activity was about 1.5-fold lower in the mutant, the substantial enzyme activity remained, suggesting the presence of a second GAPDH which is sufficient to ensure growth. The GapR protein, overproduced in Escherichia coli, was shown to bind upstream of the gap-P promoter region. The results indicate a direct role of GapR in regulation of gap expression in S. aureofaciens.
编码甘油醛-3-磷酸脱氢酶(GAPDH)的缺口基因的表达受到发育调控,并在金黄色链霉菌中由葡萄糖诱导。在缺口基因上游鉴定出一个编码与细菌转录调节因子的AraC/XylS家族相似的蛋白质的基因gapR。在分化过程中,gapR基因由单个启动子组成型表达。通过双交换的整合转化,获得了gapR基因的稳定缺失突变体。该突变仅对生长有轻微影响,对金黄色链霉菌的分化没有影响。然而,无论使用何种碳源,在金黄色链霉菌DeltagapR缺失突变体中,编码GAPDH的缺口基因的转录都大幅减少。尽管突变体中的GAPDH活性低约1.5倍,但仍保留了大量酶活性,这表明存在第二种足以确保生长的GAPDH。在大肠杆菌中过量产生的GapR蛋白显示出与缺口-P启动子区域上游结合。结果表明GapR在金黄色链霉菌中缺口基因表达的调控中起直接作用。