Ueki T, Inouye S
Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
Genes Cells. 1998 Jun;3(6):371-85. doi: 10.1046/j.1365-2443.1998.00197.x.
Myxococcus xanthus is a gram-negative bacterium that undergoes spectacular development to form multicellular fruiting bodies under nutrient deprivation. Inside a fruiting body, vegetative cells differentiate into spores. A number of sigma factors have been shown to play roles in the regulation of gene expression in the M. xanthus life cycle. Additional sigma factors were searched to further explore the M. xanthus life cycle.
A new sigma factor was identified, SigD, which consists of 297 amino acid residues. Two transcription initiation sites for the sigD gene were detected by primer extension analysis using total RNA from the vegetative and developmental cells, one of which was specific for development. The characterization of sigD-lacZ fusion strains demonstrated that sigD expression increased during entry into stationary phase of vegetative growth and during early development. A deletion mutant of sigD exhibited growth defects during the late-log phase and stationary phase, with dramatically reduced cell viability. The patterns of protein synthesis at late log phase of vegetative growth and at early development on CF agar plates were significantly different between the deletion mutant and the wild-type strain. The deletion mutant was more sensitive to various stresses when compared with the wild-type strain and did not accumulate trehalose in response to osmotic stress. The deletion mutant also showed a significant delay in fruiting body formation and sporulation and yielded fewer spores than the wild-type strain.
SigD shows characteristic features of the stationary phase sigma factors and also plays important roles in multicellular differentiation of M. xanthus.
黄色粘球菌是一种革兰氏阴性菌,在营养缺乏时会经历显著的发育过程,形成多细胞子实体。在子实体内,营养细胞分化为孢子。已证明一些σ因子在黄色粘球菌生命周期中的基因表达调控中发挥作用。为了进一步探索黄色粘球菌的生命周期,我们寻找了其他的σ因子。
我们鉴定出一个新的σ因子SigD,它由297个氨基酸残基组成。通过使用来自营养细胞和发育细胞的总RNA进行引物延伸分析,检测到sigD基因的两个转录起始位点,其中一个位点是发育特异性的。sigD-lacZ融合菌株的特性表明,sigD的表达在营养生长进入稳定期和早期发育过程中增加。sigD的缺失突变体在对数后期和稳定期表现出生长缺陷,细胞活力显著降低。在营养生长对数后期和CF琼脂平板上早期发育阶段,缺失突变体和野生型菌株的蛋白质合成模式存在显著差异。与野生型菌株相比,缺失突变体对各种胁迫更敏感,并且在渗透胁迫下不积累海藻糖。缺失突变体在子实体形成和孢子形成方面也表现出明显延迟,产生的孢子比野生型菌株少。
SigD显示出稳定期σ因子的特征,并且在黄色粘球菌的多细胞分化中也发挥重要作用。