Distinct ubiquitin ligases act sequentially for RNA polymerase II polyubiquitylation

Michelle Harreman, Michael Taschner, Stefan Sigurdsson, Roy Anindya, James Reid, Baggavalli Somesh, Stephanie E. Kong, Charles A.S. Banks, Ronald C. Conaway, Joan W. Conaway, Jesper Q. Svejstrup

Research output: Contribution to journalArticlepeer-review

114 Scopus citations

Abstract

The proteasome degrades proteins modified by polyubiquitylation, so correctly controlled ubiquitylation is crucial to avoid unscheduled proteolysis of essential proteins. The mechanism regulating proteolysis of RNAPII has been controversial since two distinct ubiquitin ligases (E3s), Rsp5 (and its human homologue NEDD4) and Elongin-Cullin complex, have both been shown to be required for its DNA-damage-induced polyubiquitylation. Here we show that these E3s work sequentially in a two-step mechanism. First, Rsp5 adds mono-ubiquitin, or sometimes a ubiquitin chain linked via ubiquitin lysine 63 that does not trigger proteolysis. When produced, the K63 chain can be trimmed to mono-ubiquitylation by an Rsp5-associated ubiquitin protease, Ubp2. Based on this mono-ubiquitin moiety on RNAPII, an Elc1/Cul3 complex then produces a ubiquitin chain linked via lysine 48, which can trigger proteolysis. Likewise, for correct polyubiquitylation of human RNAPII, NEDD4 cooperates with the ElonginA/B/C-Cullin 5 complex. These data indicate that RNAPII polyubiquitylation requires cooperation between distinct, sequentially acting ubiquitin ligases, and raise the intriguing possibility that other members of the large and functionally diverse family of NEDD4-like ubiquitin ligases also require the assistance of a second E3 when targeting proteins for degradation.

Original languageEnglish (US)
Pages (from-to)20705-20710
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number49
DOIs
StatePublished - Dec 8 2009
Externally publishedYes

Keywords

  • Elongin
  • NEDD4
  • Rsp5
  • Ubiquitylation

ASJC Scopus subject areas

  • General

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