Porter Stephanie E, Washburn Taylor M, Chang Meiping, Jaehning Judith A
Department of Biochemistry and Molecular Genetics and Molecular Biology Program, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
Eukaryot Cell. 2002 Oct;1(5):830-42. doi: 10.1128/EC.1.5.830-842.2002.
We have previously described an alternative form of RNA polymerase II in yeast lacking the Srb and Med proteins but including Pafl, Cdc73, Hprl, and Ccr4. The Pafl-RNA polymerase II complex (Paf1 complex) acts in the same pathway as the Pkc1-mitogen-activated protein kinase cascade and is required for full expression of many cell wall biosynthetic genes. The expression of several of these cell integrity genes, as well as many other Paf1-requiring genes identified by differential display and microarray analyses, is regulated during the cell cycle. To determine whether the Paf1 complex is required for basal or cyclic expression of these genes, we assayed transcript abundance throughout the cell cycle. We found that transcript abundance for a subset of cell cycle-regulated genes, including CLN1, HO, RNR1, and FAR1, is reduced from 2- to 13-fold in a paf1delta strain, but that this reduction is not promoter dependent. Despite the decreased expression levels, cyclic expression is still observed. We also examined the possibility that the Paf1 complex acts in the same pathway as either SBF (Swi4/Swi6) or MBF (Mbp1/Swi6), the partially redundant cell cycle transcription factors. Consistent with the possibility that they have overlapping essential functions, we found that loss of Paf1 is lethal in combination with loss of Swi4 or Swi6. In addition, overexpression of either Swi4 or Mbp1 suppresses some paf1delta phenotypes. These data establish that the Paf1 complex plays an important role in the essential regulatory pathway controlled by SBF and MBF.
我们之前曾描述过酵母中一种替代形式的RNA聚合酶II,它缺乏Srb和Med蛋白,但包含Pafl、Cdc73、Hprl和Ccr4。Pafl - RNA聚合酶II复合物(Paf1复合物)与Pkc1 - 丝裂原活化蛋白激酶级联反应在同一途径中起作用,并且是许多细胞壁生物合成基因充分表达所必需的。这些细胞完整性基因中的几个,以及通过差异显示和微阵列分析鉴定出的许多其他需要Paf1的基因,其表达在细胞周期中受到调控。为了确定Paf1复合物对于这些基因的基础表达或周期性表达是否必需,我们在整个细胞周期中检测了转录本丰度。我们发现,在paf1delta菌株中,包括CLN1、HO、RNR1和FAR1在内的一部分细胞周期调控基因的转录本丰度降低了2至13倍,但这种降低不依赖于启动子。尽管表达水平降低,但仍观察到周期性表达。我们还研究了Paf1复合物是否与SBF(Swi4/Swi6)或MBF(Mbp1/Swi6)这两种部分冗余的细胞周期转录因子在同一途径中起作用的可能性。鉴于它们可能具有重叠的基本功能,我们发现Paf1缺失与Swi4或Swi6缺失相结合是致死的。此外,Swi4或Mbp1的过表达可抑制一些paf1delta表型。这些数据表明,Paf1复合物在由SBF和MBF控制的基本调控途径中起重要作用。