Klein J, Grummt I
Division of Molecular Biology of the Cell II, German Cancer Research Center, D-69120 Heidelberg, Germany.
Proc Natl Acad Sci U S A. 1999 May 25;96(11):6096-101. doi: 10.1073/pnas.96.11.6096.
Transcription of ribosomal RNA genes by RNA polymerase (pol) I oscillates during the cell cycle, being maximal in S and G2 phase, repressed during mitosis, and gradually recovering during G1 progression. We have shown that transcription initiation factor (TIF)-IB/SL1 is inactivated during mitosis by cdc2/cyclin B-directed phosphorylation of TAFI110. In this study, we have monitored reactivation of transcription after exit from mitosis. We demonstrate that the pol I factor UBF is also inactivated by phosphorylation but recovers with different kinetics than TIF-IB/SL1. Whereas TIF-IB/SL1 activity is rapidly regained on entry into G1, UBF is reactivated later in G1, concomitant with the onset of pol I transcription. Repression of pol I transcription in mitosis and early G1 can be reproduced with either extracts from cells synchronized in M or G1 phase or with purified TIF-IB/SL1 and UBF isolated in the presence of phosphatase inhibitors. The results suggest that two basal transcription factors, e.g., TIF-IB/SL1 and UBF, are inactivated at mitosis and reactivated by dephosphorylation at the exit from mitosis and during G1 progression, respectively.
RNA聚合酶(pol)I对核糖体RNA基因的转录在细胞周期中呈振荡变化,在S期和G2期达到最大值,在有丝分裂期间受到抑制,并在G1期进程中逐渐恢复。我们已经表明,转录起始因子(TIF)-IB/SL1在有丝分裂期间通过TAFI110的cdc2/细胞周期蛋白B定向磷酸化而失活。在本研究中,我们监测了有丝分裂退出后转录的重新激活。我们证明,pol I因子UBF也通过磷酸化而失活,但恢复的动力学与TIF-IB/SL1不同。当进入G1期时TIF-IB/SL1活性迅速恢复,而UBF在G1期后期重新激活,与pol I转录的开始同步。有丝分裂和G1早期pol I转录的抑制可以用M期或G1期同步化细胞的提取物,或用在磷酸酶抑制剂存在下分离的纯化TIF-IB/SL1和UBF来重现。结果表明,两个基础转录因子,即TIF-IB/SL1和UBF,在有丝分裂时失活,并分别在有丝分裂退出时和G1期进程中通过去磷酸化而重新激活。