DOI: 10.1073/pnas.94.8.3656. The large subunit of RNA polymerase II is a substrate of the Rsp5 ubiquitin-protein ligase

The large subunit of RNA polymerase II is a substrate of the Rsp5 ubiquitin-protein ligase

10.1073/pnas.94.8.3656

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The E3 ubiquitin-protein ligases play an important role in controlling substrate specificity of the ubiquitin proteolysis system. A biochemical approach was taken to identify substrates of Rsp5, an essential hect (homologous to E6-AP carboxyl terminus) E3 of Saccharomyces cerevisiae . We show here that Rsp5 binds and ubiquitinates the largest subunit of RNA polymerase II (Rpb1) in vitro . Stable complex formation between Rsp5 and Rpb1 was also detected in yeast cell extracts, and repression of RSP5 expression in vivo led to an elevated steady-state level of Rpb1. The amino-terminal domain of Rsp5 mediates binding to Rpb1, while the carboxyl-terminal domain of Rpb1, containing the heptapeptide repeats characteristic of polymerase II, is necessary and sufficient for binding to Rsp5. Fusion of the Rpb1 carboxyl-terminal domain to another protein also causes that protein to be ubiquitinated by Rsp5. These findings indicate that Rsp5 targets at least a subset of cellular Rpb1 molecules for ubiquitin-dependent degradation and may therefore play a role in regulating polymerase II activities. In addition, the results support a model for hect E3 function in which the amino-terminal domain mediates substrate binding, while the carboxyl-terminal hect domain catalyzes ubiquitination of bound substrates.

Bibliography

Huibregtse, J. M., Yang, J. C., & Beaudenon, S. L. (1997). The large subunit of RNA polymerase II is a substrate of the Rsp5 ubiquitin-proteinâligase. Proceedings of the National Academy of Sciences, 94(8), 3656â3661.

Authors 3

Jon M. Huibregtse (first)
Joyce C. Yang (additional)
Sylvie L. Beaudenon (additional)

References 28 Referenced 178

10.1016/0092-8674(94)90396-4
10.1016/S0092-8674(00)81058-2
10.1128/mcb.13.2.775-784.1993
J M Huibregtse, M Scheffner, P M Howley Mol Cell Biol 13, 4918–4927 (1993). / Mol Cell Biol by Huibregtse J M (1993)
10.1016/0092-8674(93)90384-3
10.1038/373081a0
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10.1002/j.1460-2075.1990.tb07516.x
10.1016/0092-8674(95)90338-0
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10.1111/j.1365-2958.1995.mmi_18010077.x
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10.1016/0092-8674(91)90556-E

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:35 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 3:41 p.m.)
Indexed	1 month, 3 weeks ago (July 7, 2025, 3:26 a.m.)
Issued	28 years, 4 months ago (April 15, 1997)
Published	28 years, 4 months ago (April 15, 1997)
Published Online	28 years, 4 months ago (April 15, 1997)
Published Print	28 years, 4 months ago (April 15, 1997)

Funders 0

None

BibTeX

@article{Huibregtse_1997, title={The large subunit of RNA polymerase II is a substrate of the Rsp5 ubiquitin-protein ligase}, volume={94}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.94.8.3656}, DOI={10.1073/pnas.94.8.3656}, number={8}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Huibregtse, Jon M. and Yang, Joyce C. and Beaudenon, Sylvie L.}, year={1997}, month=apr, pages={3656–3661} }

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