Functional dissection of a HECT ubiquitin E3 ligase

Jin Ying Lu, Yu Yi Lin, Jiang Qian, Sheng Ce Tao, Jian Zhu, Cecile Pickart, Heng Zhu

Research output: Contribution to journalArticle

Abstract

Ubiquitination is one of the most prevalent protein post-translational modifications in eukaryotes, and its malfunction is associated with a variety of human diseases. Despite the significance of this process, the molecular mechanisms that govern the regulation of ubiquitination remain largely unknown. Here we used a combination of yeast proteome chip assays, genetic screening, and in vitro/in vivo biochemical analyses to identify and characterize eight novel in vivo substrates of the ubiquitinating enzyme Rsp5, a homolog of the human ubiquitin-ligating enzyme Nedd4, in yeast. Our analysis of the effects of a deubiquitinating enzyme, Ubp2, demonstrated that an accumulation of Lys-63-linked polyubiquitin chains results in processed forms of two substrates, Sla1 and Ygr068c. Finally we showed that the localization of another newly identified substrate, Rnr2, is Rsp5-dependent. We believe that our approach constitutes a paradigm for the functional dissection of an enzyme with pleiotropic effects.

Original languageEnglish (US)
Pages (from-to)35-45
Number of pages11
JournalMolecular and Cellular Proteomics
Volume7
Issue number1
DOIs
StatePublished - Jan 1 2008

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ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

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