Baetz K, Moffat J, Haynes J, Chang M, Andrews B
Department of Medical Genetics and Microbiology, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
Mol Cell Biol. 2001 Oct;21(19):6515-28. doi: 10.1128/MCB.21.19.6515-6528.2001.
In Saccharomyces cerevisiae, the heterodimeric transcription factor SBF (for SCB binding factor) is composed of Swi4 and Swi6 and activates gene expression at the G(1)/S-phase transition of the mitotic cell cycle. Cell cycle commitment is associated not only with major alterations in gene expression but also with highly polarized cell growth; the mitogen-activated protein kinase (MAPK) Slt2 is required to maintain cell wall integrity during periods of polarized growth and cell wall stress. We describe experiments aimed at defining the regulatory pathway involving the cell cycle transcription factor SBF and Slt2-MAPK. Gene expression assays and chromatin immunoprecipitation experiments revealed Slt2-dependent recruitment of SBF to the promoters of the G(1) cyclins PCL1 and PCL2 after activation of the Slt2-MAPK pathway. We performed DNA microarray analysis and identified other genes whose expression was reduced in both SLT2 and SWI4 deletion strains. Genes that are sensitive to both Slt2 and Swi4 appear to be uniquely regulated and reveal a role for Swi4, the DNA-binding component of SBF, which is independent of the regulatory subunit Swi6. Some of the Swi4- and Slt2-dependent genes do not require Swi6 for either their expression or for Swi4 localization to their promoters. Consistent with these results, we found a direct interaction between Swi4 and Slt2. Our results establish a new Slt2-dependent mode of Swi4 regulation and suggest roles for Swi4 beyond its prominent role in controlling cell cycle transcription.
在酿酒酵母中,异二聚体转录因子SBF(SCB结合因子)由Swi4和Swi6组成,在有丝分裂细胞周期的G(1)/S期转换时激活基因表达。细胞周期进程不仅与基因表达的重大变化有关,还与高度极化的细胞生长有关;丝裂原活化蛋白激酶(MAPK)Slt2在极化生长和细胞壁应激期间维持细胞壁完整性是必需的。我们描述了旨在确定涉及细胞周期转录因子SBF和Slt2-MAPK的调控途径的实验。基因表达分析和染色质免疫沉淀实验表明,在Slt2-MAPK途径激活后,SBF会在Slt2的作用下被招募到G(1)细胞周期蛋白PCL1和PCL2的启动子区域。我们进行了DNA微阵列分析,并鉴定出在SLT2和SWI4缺失菌株中表达均降低的其他基因。对Slt2和Swi4均敏感的基因似乎受到独特的调控,并揭示了SBF的DNA结合成分Swi4的作用,该作用独立于调节亚基Swi6。一些依赖Swi4和Slt2的基因在表达或Swi4定位到其启动子区域时都不需要Swi6。与这些结果一致,我们发现Swi4和Slt2之间存在直接相互作用。我们的结果建立了一种新的Slt2依赖的Swi4调控模式,并暗示了Swi4在控制细胞周期转录的突出作用之外的其他作用。