High-throughput sequencing reveals suppressors of Vibrio cholerae rpoE mutations: one fewer porin is enough.
High-throughput sequencing reveals suppressors of Vibrio cholerae rpoE mutations: one fewer porin is enough.
复制标题
DOI：
10.1093/nar/gkp568
复制
发表时间：
2009-09
影响因子：
14.9
通讯作者：
Waldor MK
中科院分区：
文献类型：
--
作者：
Davis BM;Waldor MK
抑制基因突变的分析对于理解基因功能是非常有价值的。然而，定位抑制基因突变的技术并不适用于大多数细菌物种。在这里，我们使用高通量测序技术来鉴定自发产生的抑制突变，这些突变能够破坏霍乱弧菌（霍乱的病原体）中的rpoE（编码σE）。由包膜应激激活的替代σ因子σE促进有助于保持和/或恢复细胞包膜完整性的因子的表达。在大肠杆菌中，rpoE是一种必需基因，只有在存在额外的抑制突变时才能被破坏。在一组独立的霍乱弧菌rpoE突变体中，超过75%含有抑制突变，其减少霍乱弧菌主要外膜孔蛋白OmpU的产生。OmpU似乎是霍乱弧菌需要和产生σE的关键决定因素。这种对单一因素的依赖性与E.在大肠杆菌中，许多因素都有助于其激活，没有一个是占主导地位的。我们还发现了一种抑制突变，它与以前描述的所有抑制基因不同，因为它提高而不是降低了σE的活性。最后，对一组rpoE突变体的分析揭示了霍乱弧菌中可能出现抑制突变的机制。
Analyses of suppressor mutations have been extremely valuable in understanding gene function. However, techniques for mapping suppressor mutations are not available for most bacterial species. Here, we used high-throughput sequencing technology to identify spontaneously arising suppressor mutations that enabled disruption of rpoE (which encodes σE) in Vibrio cholerae, the agent of cholera. The alternative sigma factor σE, which is activated by envelope stress, promotes expression of factors that help preserve and/or restore cell envelope integrity. In Escherichia coli, rpoE is an essential gene that can only be disrupted in the presence of additional suppressor mutations. Among a panel of independent V. cholerae rpoE mutants, more than 75% contain suppressor mutations that reduce production of OmpU, V. cholerae’s principal outer membrane porin. OmpU appears to be a key determinant of V. cholerae’s requirement for and production of σE. Such dependence upon a single factor contrasts markedly with regulation of σE in E. coli, in which numerous factors contribute to its activation and none is dominant. We also identified a suppressor mutation that differs from all previously described suppressors in that it elevates, rather than reduces, σE’s activity. Finally, analyses of a panel of rpoE mutants shed light on the mechanisms by which suppressor mutations may arise in V. cholerae.
登录
查看更多内容
影响因子：
3.2
作者：
Beck, BJ;Connolly, LE;Downs, DM
通讯作者：
Downs, DM
影响因子：
3.6
作者：
Mathur, Jyoti;Davis, Brigid M.;Waldor, Matthew K.
通讯作者：
Waldor, Matthew K.
影响因子：
5.6
作者：
Johansen, Jesper;Rasmussen, Anders Aamann;Valentin-Hansen, Poul
通讯作者：
Valentin-Hansen, Poul
影响因子：
3.2
作者：
Chakrabarti, SR;Chaudhuri, K;Das, J
通讯作者：
Das, J
影响因子：
3.6
作者：
Davis, Brigid M.;Waldor, Matthew K.
通讯作者：
Waldor, Matthew K.