Rao N N, Liu S, Kornberg A
Department of Biochemistry, Stanford University School of Medicine, California 94305-5307, USA.
J Bacteriol. 1998 Apr;180(8):2186-93. doi: 10.1128/JB.180.8.2186-2193.1998.
Escherichia coli transiently accumulates large amounts of inorganic polyphosphate (polyP), up to 20 mM in phosphate residues (Pi), in media deficient in both Pi and amino acids. This transient accumulation is preceded by the appearance of nucleotides ppGpp and pppGpp, generated in response to nutritional stresses. Mutants which lack PhoB, the response regulator of the phosphate regulon, do not accumulate polyP even though they develop wild-type levels of (p)ppGpp when subjected to amino acid starvation. When complemented with a phoB-containing plasmid, phoB mutants regain the ability to accumulate polyP. PolyP accumulation requires high levels of (p)ppGpp independent of whether they are generated by RelA (active during the stringent response) or SpoT (expressed during Pi starvation). Hence, accumulation of polyP requires a functional phoB gene and elevated levels of (p)ppGpp. A rapid assay of polyP depends on its adsorption to an anion-exchange disk on which it is hydrolyzed by a yeast exopolyphosphatase.
在缺乏磷酸盐(Pi)和氨基酸的培养基中,大肠杆菌会短暂积累大量无机多聚磷酸盐(polyP),其磷酸盐残基(Pi)含量高达20 mM。这种短暂积累之前会出现核苷酸ppGpp和pppGpp,它们是对营养应激作出反应而产生的。缺乏磷酸盐调节子的应答调节因子PhoB的突变体,即使在氨基酸饥饿时产生野生型水平的(p)ppGpp,也不会积累多聚磷酸盐。当用含有phoB的质粒进行互补时,phoB突变体恢复了积累多聚磷酸盐的能力。多聚磷酸盐的积累需要高水平的(p)ppGpp,无论它们是由RelA(在严格应答期间活跃)还是SpoT(在Pi饥饿期间表达)产生的。因此,多聚磷酸盐的积累需要一个功能性的phoB基因和升高的(p)ppGpp水平。多聚磷酸盐的快速检测取决于其对阴离子交换盘的吸附,在该盘上它会被酵母外切多聚磷酸酶水解。