Magill N G, Cowan A E, Leyva-Vazquez M A, Brown M, Koppel D E, Setlow P
Department of Biochemistry, University of Connecticut Health Center, Farmington 06030, USA.
J Bacteriol. 1996 Apr;178(8):2204-10. doi: 10.1128/jb.178.8.2204-2210.1996.
Analysis of the pH decrease and 3-phosphoglyceric acid (3PGA) accumulation in the forespore compartment of sporulating cells of Bacillus subtilis showed that the pH decrease of 1 to 1.2 units at approximately 4 h of sporulation preceded 3PGA accumulation, as observed previously in B. megaterium. These data, as well as analysis of the forespore pH decrease in asporogenous mutants of B. subtilis, indicated that sigma G-dependent forespore transcription, but not sigma K-dependent mother cell transcription, is required for the forespore pH decrease. Further analysis of these asporogenous mutants showed an excellent correlation between the forespore pH decrease and the forespore's accumulation of 3PGA. These latter results are consistent with our previous suggestion that the decrease in forespore pH results in greatly decreased activity of phosphoglycerate mutase in the forespore, which in turn leads to 3PGA accumulation. In further support of this suggestion, we found that (i) elevating the pH of developing forespores of B. megaterium resulted in rapid utilization of the forespore's 3PGA depot and (ii) increasing forespore levels of PGM approximately 10-fold in B. subtilis resulted in a large decrease in the spore's depot of 3PGA. The B. subtilis strain with a high phosphoglycerate mutase level sporulated, and the spores germinated and went through outgrowth normally, indicating that forespore accumulation of a large 3PGA depot is not essential for these processes.
对枯草芽孢杆菌芽孢形成细胞前芽孢区室中pH值下降和3-磷酸甘油酸(3PGA)积累的分析表明,在芽孢形成约4小时时,pH值下降1至1.2个单位先于3PGA积累,这与之前在巨大芽孢杆菌中观察到的情况一致。这些数据,以及对枯草芽孢杆菌无芽孢突变体前芽孢pH值下降的分析表明,前芽孢pH值下降需要σG依赖性前芽孢转录,而不是σK依赖性母细胞转录。对这些无芽孢突变体的进一步分析表明,前芽孢pH值下降与前芽孢中3PGA的积累之间存在极好的相关性。后一个结果与我们之前的推测一致,即前芽孢pH值的下降导致前芽孢中磷酸甘油酸变位酶的活性大大降低,进而导致3PGA积累。为进一步支持这一推测,我们发现:(i)提高巨大芽孢杆菌发育中前芽孢的pH值会导致前芽孢中3PGA储备的快速利用;(ii)在枯草芽孢杆菌中将前芽孢中PGM的水平提高约10倍会导致芽孢中3PGA储备的大幅减少。具有高磷酸甘油酸变位酶水平的枯草芽孢杆菌菌株能够形成芽孢,并且芽孢能够正常萌发和生长,这表明前芽孢中大量3PGA储备的积累对于这些过程并非必不可少。