May G, Dersch P, Haardt M, Middendorf A, Bremer E
Department of Biology, University of Konstanz, Federal Republic of Germany.
Mol Gen Genet. 1990 Oct;224(1):81-90. doi: 10.1007/BF00259454.
A class of trans-acting mutations, which alter the osmoregulated expression of the Escherichia coli proU operon, maps at 27 min on the chromosome in a locus we have called osmZ. Mutations in osmZ are allelic to bglY, pilG and virR, affect gene expression, increase the frequency of the site-specific DNA inversion mediating fimbrial phase variation, stimulate the formation of deletions, and influence in vivo supercoiling of reporter plasmids. We have cloned the osmZ+ gene, mapped it at 1307 kb of the E. coli restriction map, identified its gene product as a 16 kDa protein, and determined the nucleotide sequence of the osmZ+ gene. The deduced amino acid sequence for OsmZ predicts a protein of 137 amino acid residues with a calculated molecular weight of 15,530. The primary sequence of OsmZ is identical to that of H-NS (H1a), a DNA-binding protein that affects DNA topology and is known to be associated with the bacterial nucleoid. Thus, osmZ is the structural gene for the H-NS (H1a) protein. The nucleotide sequence of osmZ is almost identical to that of hns; however, hns was incorrectly located at 6.1 min on the E. coli linkage map. Increased osmZ gene dosage leads to cell filament formation, altered gene expression, and reduced frequency of fimbrial phase variation. Our results suggest that the nucleoid-associated DNA-binding protein H-NS (H1a) plays a critical role in gene expression and in determining the structure of the genetic material.
一类反式作用突变可改变大肠杆菌proU操纵子的渗透调节表达,这些突变位于染色体上27分钟处的一个位点,我们称之为osmZ。osmZ中的突变与bglY、pilG和virR等位,影响基因表达,增加介导菌毛相变的位点特异性DNA倒位频率,刺激缺失形成,并影响报告质粒在体内的超螺旋。我们克隆了osmZ⁺基因,将其定位在大肠杆菌限制酶图谱的1307 kb处,确定其基因产物为一种16 kDa的蛋白质,并测定了osmZ⁺基因的核苷酸序列。推导的OsmZ氨基酸序列预测该蛋白质由137个氨基酸残基组成,计算分子量为15,530。OsmZ的一级序列与H-NS(H1a)相同,H-NS是一种影响DNA拓扑结构且已知与细菌类核相关的DNA结合蛋白。因此,osmZ是H-NS(H1a)蛋白的结构基因。osmZ的核苷酸序列与hns几乎相同;然而,hns在大肠杆菌连锁图谱上的位置错误地定在6.1分钟处。增加osmZ基因剂量会导致细胞丝状化、基因表达改变以及菌毛相变频率降低。我们的结果表明,类核相关的DNA结合蛋白H-NS(H1a)在基因表达和确定遗传物质结构方面起着关键作用。