Calamita G, Kempf B, Bonhivers M, Bishai W R, Bremer E, Agre P
Dipartimento di Fisiologia Generale ed Ambientale, University of Bari, via Amendola, 165/A, 70126 Bari, Italy.
Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3627-31. doi: 10.1073/pnas.95.7.3627.
Osmotic movement of water across bacterial cell membranes is postulated to be a homeostatic mechanism for maintaining cell turgor. The molecular water transporter remained elusive until discovery of the Escherichia coli water channel, AqpZ, however the regulation of the aqpZ gene expression and physiological function of the AqpZ protein are unknown. Northern analysis revealed a transcript of 0.7 kb, confirming the monocistronic nature of aqpZ. Regulatory studies performed with an aqpZ::lacZ low copy plasmid demonstrate enhanced expression during mid-logarithmic growth, and expression of the gene is dependent upon the extracellular osmolality, which increased in hypoosmotic environments but strongly reduced in hyperosmolar NaCl or KCl. While disruption of the chromosomal aqpZ is not lethal for E. coli, the colonies of the aqpZ knockout mutant are smaller than those of the parental wild-type strain. When cocultured with parental wild-type E. coli, the aqpZ knockout mutant exhibits markedly reduced colony formation when grown at 39 degrees C. Similarly, the aqpZ knockout mutant also exhibits greatly reduced colony formation when grown at low osmolality, but this phenotype is reversed by overexpression of AqpZ protein. These results implicate AqpZ as a participant in the adaptive response of E. coli to hypoosmotic environments and indicate a requirement for AqpZ by rapidly growing cells.
水通过细菌细胞膜的渗透运动被假定为维持细胞膨压的一种稳态机制。在发现大肠杆菌水通道蛋白AqpZ之前,分子水转运体一直难以捉摸,然而,aqpZ基因表达的调控以及AqpZ蛋白的生理功能尚不清楚。Northern分析显示有一个0.7 kb的转录本,证实了aqpZ的单顺反子性质。用一个aqpZ::lacZ低拷贝质粒进行的调控研究表明,在对数生长中期表达增强,并且该基因的表达依赖于细胞外渗透压,在低渗环境中渗透压升高,但在高渗的NaCl或KCl中则大幅降低。虽然染色体上的aqpZ缺失对大肠杆菌并非致命,但aqpZ基因敲除突变体的菌落比亲本野生型菌株的菌落小。当与亲本野生型大肠杆菌共培养时,aqpZ基因敲除突变体在39℃生长时菌落形成明显减少。同样,aqpZ基因敲除突变体在低渗透压下生长时菌落形成也大大减少,但这种表型可通过AqpZ蛋白的过表达得到逆转。这些结果表明AqpZ参与了大肠杆菌对低渗环境的适应性反应,并表明快速生长的细胞需要AqpZ。