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大肠杆菌中精氨酸生物合成操纵子argCBH的稳定期表达。

Stationary phase expression of the arginine biosynthetic operon argCBH in Escherichia coli.

作者信息

Weerasinghe Jeevaka P, Dong Tao, Schertzberg Michael R, Kirchhof Mark G, Sun Yuan, Schellhorn Herb E

机构信息

McMaster University, Department of Biology, Life Sciences Building, Rm, 218, 1280 Main Street West, Hamilton, ON, Canada, L8S 4K1.

出版信息

BMC Microbiol. 2006 Feb 22;6:14. doi: 10.1186/1471-2180-6-14.

Abstract

BACKGROUND

Arginine biosynthesis in Escherichia coli is elevated in response to nutrient limitation, stress or arginine restriction. Though control of the pathway in response to arginine limitation is largely modulated by the ArgR repressor, other factors may be involved in increased stationary phase and stress expression.

RESULTS

In this study, we report that expression of the argCBH operon is induced in stationary phase cultures and is reduced in strains possessing a mutation in rpoS, which encodes an alternative sigma factor. Using strains carrying defined argR, and rpoS mutations, we evaluated the relative contributions of these two regulators to the expression of argH using operon-lacZ fusions. While ArgR was the main factor responsible for modulating expression of argCBH, RpoS was also required for full expression of this biosynthetic operon at low arginine concentrations (below 60 microM L-arginine), a level at which growth of an arginine auxotroph was limited by arginine. When the argCBH operon was fully de-repressed (arginine limited), levels of expression were only one third of those observed in deltaargR mutants, indicating that the argCBH operon is partially repressed by ArgR even in the absence of arginine. In addition, argCBH expression was 30-fold higher in deltaargR mutants relative to levels found in wild type, fully-repressed strains, and this expression was independent of RpoS.

CONCLUSION

The results of this study indicate that both derepression and positive control by RpoS are required for full control of arginine biosynthesis in stationary phase cultures of E. coli.

摘要

背景

在大肠杆菌中,精氨酸生物合成会因营养限制、应激或精氨酸限制而增加。尽管该途径对精氨酸限制的调控主要由ArgR阻遏物介导,但其他因素可能参与了稳定期和应激表达的增加。

结果

在本研究中,我们报道了argCBH操纵子在稳定期培养物中被诱导表达,而在编码替代σ因子的rpoS发生突变的菌株中表达降低。我们使用携带特定argR和rpoS突变的菌株,通过操纵子-lacZ融合来评估这两种调节因子对argH表达的相对贡献。虽然ArgR是调节argCBH表达的主要因素,但在低精氨酸浓度(低于60μM L-精氨酸)下,RpoS也是该生物合成操纵子完全表达所必需的,在这个浓度下,精氨酸营养缺陷型的生长受到精氨酸的限制。当argCBH操纵子完全去阻遏(精氨酸受限)时,表达水平仅为在deltaargR突变体中观察到的三分之一,这表明即使在没有精氨酸的情况下,argCBH操纵子也受到ArgR的部分抑制。此外,相对于野生型完全阻遏菌株,deltaargR突变体中的argCBH表达高30倍,且这种表达与RpoS无关。

结论

本研究结果表明,在大肠杆菌稳定期培养物中,RpoS的去阻遏和正调控对于精氨酸生物合成的完全控制都是必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/59f9/1413537/e4b6801da4b7/1471-2180-6-14-1.jpg

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