期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 284, 期 27, 页码 18377-18389出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M109.001503
关键词
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资金
- Biotechnology and Biological Sciences Research Council, UK
- Biotechnology and Biological Sciences Research Council [BB/C509831/1] Funding Source: researchfish
Zinc ions play indispensable roles in biological chemistry. However, bacteria have an impressive ability to acquire Zn2+ from the environment, making it exceptionally difficult to achieve Zn2+ deficiency, and so a comprehensive understanding of the importance of Zn2+ has not been attained. Reduction of the Zn2+ content of Escherichia coli growth medium to 60 nM or less is reported here for the first time, without recourse to chelators of poor specificity. Cells grown in Zn2+-deficient medium had a reduced growth rate and contained up to five times less cellular Zn2+. To understand global responses to Zn2+ deficiency, microarray analysis was conducted of cells grown under Zn2+-replete and Zn2+-depleted conditions in chemostat cultures. Nine genes were up-regulated more than 2-fold (p<0.05) in cells from Zn2+-deficient chemostats, including zinT (yodA). zinT is shown to be regulated by Zur ( zinc uptake regulator). A mutant lacking zinT displayed a growth defect and a 3-fold lowered cellular Zn2+ level under Zn2+ limitation. The purified ZinT protein possessed a single, high affinity metal-binding site that can accommodate Zn2+ or Cd2+. A further up-regulated gene, ykgM, is believed to encode a non-Zn2+ finger-containing paralogue of the Zn2+ finger ribosomal protein L31. The gene encoding the periplasmic Zn2+- binding protein znuA showed increased expression. During both batch and chemostat growth, cells found more Zn2+ than was originally added to the culture, presumably because of leaching from the culture vessel. Zn2+ elimination is shown to be a more precise method of depleting Zn2+ than by using the chelator N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine.
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