Schroeder Stephanie C, Zorio Diego A R, Schwer Beate, Shuman Stewart, Bentley David
Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, B121, 4200 East 9th Avenue, Denver, CO 80262 USA.
Mol Cell. 2004 Feb 13;13(3):377-87. doi: 10.1016/s1097-2765(04)00007-3.
Capping enzymes bind the phosphorylated pol II CTD permitting cotranscriptional capping of nascent pre-mRNAs. We asked whether these interactions influence pol II function using ChIP in ts mutants of yeast capping enzymes. Pol II occupancy at the 5' ends of PGK1, ENO2, GAL1, and GAL10 was reduced by inactivation of the methyltransferase, Abd1, but not the guanylyltransferase, Ceg1, suggesting that Abd1 contributes to stable promoter binding. At other genes, Abd1 inactivation increased the 5':3' ratio of pol II density in the promoter-proximal region and caused Ser5 hyperphosphorylation of the pol II CTD. These results suggest an additional role for Abd1 in the promoter clearance and/or promoter-proximal elongation steps of transcription. The transcriptional functions of Abd1 are independent of methyltransferase activity. Manipulation of transcription by Abd1 may enhance cotranscriptional capping and also act as a checkpoint to ensure that a nascent transcript has a cap before it can be completed.
加帽酶结合磷酸化的聚合酶II CTD,从而对新生的前体mRNA进行共转录加帽。我们利用酵母加帽酶的温度敏感型突变体,通过染色质免疫沉淀(ChIP)技术来探究这些相互作用是否会影响聚合酶II的功能。甲基转移酶Abd1失活会降低PGK1、ENO2、GAL1和GAL10基因5'端的聚合酶II占有率,但鸟苷酸转移酶Ceg1失活则不会,这表明Abd1有助于稳定启动子结合。在其他基因中,Abd1失活会增加启动子近端区域聚合酶II密度的5':3'比率,并导致聚合酶II CTD的Ser5过度磷酸化。这些结果表明Abd1在转录的启动子清除和/或启动子近端延伸步骤中具有额外作用。Abd1的转录功能独立于甲基转移酶活性。Abd1对转录的调控可能会增强共转录加帽,并且还可作为一个检查点,以确保新生转录本在完成之前具有帽子结构。