Ho Y, Costanzo M, Moore L, Kobayashi R, Andrews B J
Department of Molecular and Medical Genetics, University of Toronto, Toronto, Canada M5S 1A8.
Mol Cell Biol. 1999 Aug;19(8):5267-78. doi: 10.1128/MCB.19.8.5267.
In Saccharomyces cerevisiae, gene expression in the late G(1) phase is activated by two transcription factors, SBF and MBF. SBF contains the Swi4 and Swi6 proteins and activates the transcription of G(1) cyclin genes, cell wall biosynthesis genes, and the HO gene. MBF is composed of Mbp1 and Swi6 and activates the transcription of genes required for DNA synthesis. Mbp1 and Swi4 are the DNA binding subunits for MBF and SBF, while the common subunit, Swi6, is presumed to play a regulatory role in both complexes. We show that Stb1, a protein first identified in a two-hybrid screen with the transcriptional repressor Sin3, binds Swi6 in vitro. The STB1 transcript was cell cycle periodic and peaked in late G(1) phase. In vivo accumulation of Stb1 phosphoforms was dependent on CLN1, CLN2, and CLN3, which encode G(1)-specific cyclins for the cyclin-dependent kinase Cdc28, and Stb1 was phosphorylated by Cln-Cdc28 kinases in vitro. Deletion of STB1 caused an exacerbated delay in G(1) progression and the onset of Start transcription in a cln3Delta strain. Our results suggest a role for STB1 in controlling the timing of Start transcription that is revealed in the absence of the G(1) regulator CLN3, and they implicate Stb1 as an in vivo target of G(1)-specific cyclin-dependent kinases.
在酿酒酵母中,G1期晚期的基因表达由两种转录因子SBF和MBF激活。SBF包含Swi4和Swi6蛋白,并激活G1期细胞周期蛋白基因、细胞壁生物合成基因和HO基因的转录。MBF由Mbp1和Swi6组成,并激活DNA合成所需基因的转录。Mbp1和Swi4是MBF和SBF的DNA结合亚基,而共同亚基Swi6被认为在这两种复合物中都发挥调节作用。我们发现,Stb1是一种首先在与转录抑制因子Sin3的双杂交筛选中鉴定出的蛋白,它在体外与Swi6结合。STB1转录本呈细胞周期周期性变化,并在G1期晚期达到峰值。Stb1磷酸化形式的体内积累依赖于CLN1、CLN2和CLN3,它们编码细胞周期蛋白依赖性激酶Cdc28的G1期特异性细胞周期蛋白,并且Stb1在体外被Cln-Cdc28激酶磷酸化。在cln3Δ菌株中,STB1的缺失导致G1期进程和起始转录起始的延迟加剧。我们的结果表明,STB1在控制起始转录的时间方面发挥作用,这在缺乏G1期调节因子CLN3的情况下得以揭示,并且它们表明Stb1是G1期特异性细胞周期蛋白依赖性激酶的体内靶点。