title: Polyubiquitylation of AMF requires cooperation between the gp78 and TRIM25 ubiquitin ligases.
abstract: gp78 is a ubiquitin ligase that plays a vital role in endoplasmic reticulum (ER)-associated degradation (ERAD). Here we report that autocrine motility factor (AMF), also known as phosphoglucose isomerase (PGI), is a novel substrate of gp78. We show that polyubiquitylation of AMF requires cooperative interaction between gp78 and the ubiquitin ligase TRIM25 (tripartite motif-containing protein 25). While TRIM25 mediates the initial round of ubiquitylation, gp78 catalyzes polyubiquitylation of AMF. The E4-like activity of gp78 was illustrated by an in vitro polyubiquitylation assay using Ub-DHFR as a model substrate. We further demonstrate that TRIM25 ubiquitylates gp78 and that overexpression of TRIM25 accelerates the degradation of gp78. Our data suggest that TRIM25 not only cooperates with gp78 in polyubiquitylation of AMF but also gauges the steady-state level of gp78. This study uncovers a previously unknown functional link between gp78 and TRIM25 and provides mechanistic insight into gp78-mediated protein ubiquitylation.
authors.affiliation: Barbara Ann Karmanos Cancer Institute and Department of Oncology, Wayne State University School of Medicine, Detroit, MI, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Seung-Wook
authors.initials: SW
authors.lastname: Ha
authors.forename: Tianpeng
authors.initials: T
authors.lastname: Zhang
authors.forename: Dhong-Hyo
authors.initials: DH
authors.lastname: Kho
authors.forename: Avraham
authors.initials: A
authors.lastname: Raz
authors.forename: Youming
authors.initials: Y
authors.lastname: Xie
mesh_terms: D005956:Glucose-6-Phosphate Isomerase / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D060285:Receptors, Autocrine Motility Factor / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D014157:Transcription Factors; D000070584:Tripartite Motif Proteins; C555000:AMFR protein, human; D060285:Receptors, Autocrine Motility Factor; C084523:TRIM25 protein, human; D044767:Ubiquitin-Protein Ligases; D005956:Glucose-6-Phosphate Isomerase