DNA Damage-induced Def1-RNA Polymerase II Interaction and Def1 Requirement for Polymerase Ubiquitylation in Vitro

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DNA damage-induced Def1-RNA polymerase II interaction and Def1 requirement for polymerase ubiquitylation in vitro.

UV-induced DNA damage results in ubiquitylation and degradation of RNA polymerase II (RNAPII). In yeast, this requires the DEF1 gene, the product of which forms a complex with the transcription-coupling repair factor, Rad26. However, whether Def1 is directly involved in RNAPII ubiquitylation has remained unclear. Here we report the establishment of a reconstituted system for studying UV-induced...

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Def1 Promotes the Degradation of Pol3 for Polymerase Exchange to Occur During DNA-Damage–Induced Mutagenesis in Saccharomyces cerevisiae

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Multiple Mechanisms Confining RNA Polymerase II Ubiquitylation to Polymerases Undergoing Transcriptional Arrest

In order to study mechanisms and regulation of RNA polymerase II (RNAPII) ubiquitylation and degradation, highly purified factors were used to reconstitute RNAPII ubiquitylation in vitro. We show that arrested RNAPII elongation complexes are the preferred substrates for ubiquitylation. Accordingly, not only DNA-damage-dependent but also DNA-damage-independent transcriptional arrest results in R...

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Proteasome-Mediated Processing of Def1, a Critical Step in the Cellular Response to Transcription Stress

DNA damage triggers polyubiquitylation and degradation of the largest subunit of RNA polymerase II (RNAPII), a "mechanism of last resort" employed during transcription stress. In yeast, this process is dependent on Def1 through a previously unresolved mechanism. Here, we report that Def1 becomes activated through ubiquitylation- and proteasome-dependent processing. Def1 processing results in th...

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Transcriptional arrest triggers ubiquitylation of RNA polymerase II (RNAPII). We mapped the yeast RNAPII ubiquitylation sites and found that they play an important role in elongation and the DNA-damage response. One site lies in a protein domain that is unordered in free RNAPII, but ordered in the elongating form, helping explain the preferential ubiquitylation of this form. The other site is >...

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ژورنال

عنوان ژورنال: Journal of Biological Chemistry

سال: 2004

ISSN: 0021-9258

DOI: 10.1074/jbc.c400185200