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Molecular and Cellular Biology, October 1999, p. 6972-6979, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Rsp5 Ubiquitin-Protein Ligase Mediates DNA
Damage-Induced Degradation of the Large Subunit of RNA Polymerase
II in Saccharomyces cerevisiae
Sylvie L.
Beaudenon,1
Maria R.
Huacani,2
Guangli
Wang,1
Donald P.
McDonnell,2 and
Jon M.
Huibregtse1,*
Department of Molecular Biology and
Biochemistry, Rutgers University, Piscataway, New Jersey
08855,1 and Department of
Pharmacology and Cancer Biology, Duke University Medical Center,
Durham, North Carolina 277102
Received 26 April 1999/Returned for modification 10 June
1999/Accepted 1 July 1999
Rsp5 is an E3 ubiquitin-protein ligase of Saccharomyces
cerevisiae that belongs to the hect domain family of E3 proteins. We have previously shown that Rsp5 binds and ubiquitinates the largest
subunit of RNA polymerase II, Rpb1, in vitro. We show here that Rpb1
ubiquitination and degradation are induced in vivo by UV irradiation
and by the UV-mimetic compound 4-nitroquinoline-1-oxide (4-NQO) and
that a functional RSP5 gene product is required for this
effect. The 26S proteasome is also required; a mutation of SEN3/RPN2 (sen3-1), which encodes an essential
regulatory subunit of the 26S proteasome, partially blocks
4-NQO-induced degradation of Rpb1. These results suggest that
Rsp5-mediated ubiquitination and degradation of Rpb1 are components of
the response to DNA damage. A human WW domain-containing hect (WW-hect)
E3 protein closely related to Rsp5, Rpf1/hNedd4, also binds and
ubiquitinates both yeast and human Rpb1 in vitro, suggesting that Rpf1
and/or another WW-hect E3 protein mediates UV-induced degradation of the large subunit of polymerase II in human cells.
*
Corresponding author. Mailing address: Department of
Molecular Biology and Biochemistry, Rutgers University, Picataway, NJ 08855. Phone: (732) 445-0938. Fax: (732) 445-4213. E-mail:
huibregt{at}waksman.rutgers.edu.
Molecular and Cellular Biology, October 1999, p. 6972-6979, Vol. 19, No. 10
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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