Rpb1 Sumoylation in Response to UV Radiation or Transcriptional Impairment in Yeast

Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, United States of America.
PLoS ONE (Impact Factor: 3.23). 02/2009; 4(4):e5267. DOI: 10.1371/journal.pone.0005267
Source: PubMed


Covalent modifications of proteins by ubiquitin and the Small Ubiquitin-like MOdifier (SUMO) have been revealed to be involved in a plethora of cellular processes, including transcription, DNA repair and DNA damage responses. It has been well known that in response to DNA damage that blocks transcription elongation, Rpb1, the largest subunit of RNA polymerase II (Pol II), is ubiquitylated and subsequently degraded in mammalian and yeast cells. However, it is still an enigma regarding how Pol II responds to damaged DNA and conveys signal(s) for DNA damage-related cellular processes. We found that Rpb1 is also sumoylated in yeast cells upon UV radiation or impairment of transcription elongation, and this modification is independent of DNA damage checkpoint activation. Ubc9, an E2 SUMO conjugase, and Siz1, an E3 SUMO ligase, play important roles in Rpb1 sumoylation. K1487, which is located in the acidic linker region between the C-terminal domain and the globular domain of Rpb1, is the major sumoylation site. Rpb1 sumoylation is not affected by its ubiquitylation, and vice versa, indicating that the two processes do not crosstalk. Abolishment of Rpb1 sumoylation at K1487 does not affect transcription elongation or transcription coupled repair (TCR) of UV-induced DNA damage. However, deficiency in TCR enhances UV-induced Rpb1 sumoylation, presumably due to the persistence of transcription-blocking DNA lesions in the transcribed strand of a gene. Remarkably, abolishment of Rpb1 sumoylation at K1487 causes enhanced and prolonged UV-induced phosphorylation of Rad53, especially in TCR-deficient cells, suggesting that the sumoylation plays a role in restraining the DNA damage checkpoint response caused by transcription-blocking lesions. Our results demonstrate a novel covalent modification of Rpb1 in response to UV induced DNA damage or transcriptional impairment, and unravel an important link between the modification and the DNA damage checkpoint response.

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    • "These include multiple subunits of the general transcription factors (GTFs) TFIIA, TFIIF, and TFIID (TBP, as well as several TAFs); Mediator; and subunits of RNA polymerase II (RNAP II) itself. Of the multiple yeast subunits of RNAP II found to be sumoylated, Rpb1 sumoylation was characterized and appears to occur as part of the UV response, but blocking Rpb1 sumoylation does not affect transcription elongation or cell growth (Chen et al. 2009). Sumoylation of the human TFIID subunit TAF5 was shown to impair TFIID binding to promoter DNA (Boyer-Guittaut et al. 2005), suggesting that sumoylation might be a negative regulator of TFIID as well. "
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