BRCA1/BARD1 inhibition of mRNA 3′ processing involves targeted degradation of RNA polymerase II
- Frida E. Kleiman1,2,
- Foon Wu-Baer3,
- Danae Fonseca2,
- Syuzo Kaneko1,
- Richard Baer3, and
- James L. Manley1,4
- 1Department of Biological Sciences, Columbia University, New York, New York 10027, USA; 2Chemistry Department, Hunter College, City University of New York, New York, New York 10021, USA; 3Department of Pathology, Institute for Cancer Genetics, Medical Center, Columbia University, New York, New York 10032, USA
Abstract
Mammalian cells exhibit a complex response to DNA damage. The tumor suppressor BRCA1 and associated protein BARD1 are thought to play an important role in this response, and our previous work demonstrated that this includes transient inhibition of the pre-mRNA 3′ processing machinery. Here we provide evidence that this inhibition involves proteasomal degradation of a component necessary for processing, RNA polymerase II (RNAP II). We further show that RNAP IIO, the elongating form of the enzyme, is a specific in vitro target of the BRCA1/BARD1 ubiquitin ligase activity. Significantly, siRNA-mediated knockdown of BRCA1 and BARD1 resulted in stabilization of RNAP II after DNA damage. In addition, inhibition of 3′ cleavage induced by DNA damage was reverted in extracts of BRCA1-, BARD1-, or BRCA1/BARD1-depleted cells. We also describe corresponding changes in the nuclear localization and/or accumulation of these factors following DNA damage. Our results support a model in which a BRCA1/BARD1-containing complex functions to initiate degradation of stalled RNAP IIO, inhibiting the coupled transcription-RNA processing machinery and facilitating repair.
