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  • A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation.

A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation.

Nature communications (2014-05-09)
Michael L van de Weijer, Michael C Bassik, Rutger D Luteijn, Cornelia M Voorburg, Mirjam A M Lohuis, Elisabeth Kremmer, Rob C Hoeben, Emily M LeProust, Siyuan Chen, Hanneke Hoelen, Maaike E Ressing, Weronika Patena, Jonathan S Weissman, Michael T McManus, Emmanuel J H J Wiertz, Robert Jan Lebbink
ABSTRACT

Misfolded ER proteins are retrotranslocated into the cytosol for degradation via the ubiquitin-proteasome system. The human cytomegalovirus protein US11 exploits this ER-associated protein degradation (ERAD) pathway to downregulate HLA class I molecules in virus-infected cells, thereby evading elimination by cytotoxic T-lymphocytes. US11-mediated degradation of HLA class I has been instrumental in the identification of key components of mammalian ERAD, including Derlin-1, p97, VIMP and SEL1L. Despite this, the process governing retrotranslocation of the substrate is still poorly understood. Here using a high-coverage genome-wide shRNA library, we identify the uncharacterized protein TMEM129 and the ubiquitin-conjugating E2 enzyme UBE2J2 to be essential for US11-mediated HLA class I downregulation. TMEM129 is an unconventional C4C4-type RING finger E3 ubiquitin ligase that resides within a complex containing various other ERAD components, including Derlin-1, Derlin-2, VIMP and p97, indicating that TMEM129 is an integral part of the ER-resident dislocation complex mediating US11-induced HLA class I degradation.

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