Transcription-coupled and DNA damage-dependent ubiquitination of RNA polymerase II in vitro
- 19 March 2002
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 99 (7), 4239-4244
- https://doi.org/10.1073/pnas.072068399
Abstract
Transcription-coupled repair (TCR) is essential for the rapid, preferential removal of DNA damage in active genes. The large subunit of RNA polymerase (Pol) II is ubiquitinated in cells after UV-irradiation or cisplatin treatment, which induces DNA damage preferentially repaired by TCR. Several human mutations, such as Cockayne syndrome complementation groups A and B, are defective in TCR and incapable of Pol II ubiquitination upon DNA damage. Here we demonstrate a correlation between ubiquitination of RNA Pol II and arrest of transcription in vitro. Ubiquitination of Pol II is significantly induced by α-amanitin, an amatoxin that blocks Pol II elongation and causes its degradation in cells. Pol II undergoes similar ubiquitination on DNA containing cisplatin adducts that arrest transcription. Stimulation of ubiquitination requires the addition of template DNA and does not occur in the presence of an antibody to the general transcription factor TFIIB, indicating the transcription dependence of the reaction. We propose that components of the reaction recognize elongating Pol II–DNA complexes arrested by α-amanitin or cisplatin lesions, triggering ubiquitination.Keywords
This publication has 73 references indexed in Scilit:
- RETRACTED: Transcription-Coupled Repair of 8-oxoGuanine: Requirement for XPG, TFIIH, and CSB and Implications for Cockayne SyndromeCell, 2000
- p53 inhibits entry into mitosis when DNA synthesis is blockedOncogene, 1999
- Ultraviolet Radiation-induced Ubiquitination and Proteasomal Degradation of the Large Subunit of RNA Polymerase IIJournal of Biological Chemistry, 1998
- Identification of a RING protein that can interact in vivo with the BRCA1 gene productNature Genetics, 1996
- The HMG-domain protein Ixr1 blocks excision repair of cisplatin-DNA adducts in yeastMutation Research/DNA Repair, 1996
- DNA Adducts of cis-Diamminedichloroplatinum(II) and Its Trans Isomer Inhibit RNA Polymerase II Differentially in VivoBiochemistry, 1995
- Spectrum of DNA-platinum adduct recognition by prokaryotic and eukaryotic DNA-dependent RNA polymerasesBiochemistry, 1993
- Promoter specificity of basal transcription factorsCell, 1992
- Formation of a single phosphodiester bond by RNA polymerase B from calf thymus is not inhibited by .alpha.-amanitinBiochemistry, 1982
- α-Amanitin: A specific inhibitor of one of two DNA-dependent RNA polymerase activities from calf thymusBiochemical and Biophysical Research Communications, 1970