Rosario Christopher J, Singer Mitchell
Section of Microbiology, The University of California, Davis, Davis, CA 95616, USA.
J Bacteriol. 2007 Dec;189(24):8793-800. doi: 10.1128/JB.01361-07. Epub 2007 Sep 28.
Under conditions of nutrient deprivation, Myxococcus xanthus undergoes a developmental process that results in the formation of a fruiting body containing environmentally resistant myxospores. We have shown that myxospores contain two copies of the genome, suggesting that cells must replicate the genome prior to or during development. To further investigate the role of DNA replication in development, a temperature-sensitive dnaB mutant, DnaB(A116V), was isolated from M. xanthus. Unlike what happens in Escherichia coli dnaB mutants, where DNA replication immediately halts upon a shift to a nonpermissive temperature, growth and DNA replication of the M. xanthus mutant ceased after one cell doubling at a nonpermissive temperature, 37 degrees C. We demonstrated that at the nonpermissive temperature the DnaB(A116V) mutant arrested as a population of 1n cells, implying that these cells could complete one round of the cell cycle but did not initiate new rounds of DNA replication. In developmental assays, the DnaB(A116V) mutant was unable to develop into fruiting bodies and produced fewer myxospores than the wild type at the nonpermissive temperature. However, the mutant was able to undergo development when it was shifted to a permissive temperature, suggesting that cells had the capacity to undergo DNA replication during development and to allow the formation of myxospores.
在营养剥夺条件下,黄色黏球菌会经历一个发育过程,最终形成一个含有抗环境压力的黏孢子的子实体。我们已经表明,黏孢子含有两份基因组,这表明细胞在发育之前或发育过程中必须复制基因组。为了进一步研究DNA复制在发育中的作用,从黄色黏球菌中分离出了一种温度敏感型dnaB突变体DnaB(A116V)。与大肠杆菌dnaB突变体不同,在大肠杆菌中,当转移到非允许温度时DNA复制会立即停止,而黄色黏球菌突变体在37℃的非允许温度下经过一次细胞倍增后,其生长和DNA复制就停止了。我们证明,在非允许温度下,DnaB(A116V)突变体停滞为一群1n细胞,这意味着这些细胞可以完成一轮细胞周期,但不会启动新一轮的DNA复制。在发育试验中,DnaB(A116V)突变体在非允许温度下无法发育成子实体,并且产生的黏孢子比野生型少。然而,当突变体转移到允许温度时,它能够进行发育,这表明细胞在发育过程中有能力进行DNA复制并形成黏孢子。