Membrane-Associated RING-CH proteins associate with Bap31 and target CD81 and CD44 to lysosomes

• Published in: PloS one Vol. 5; no. 12; p. e15132
• Main Authors: Bartee, Eric, Eyster, Craig A, Viswanathan, Kasinath, Mansouri, Mandana, Donaldson, Julie G, Früh, Klaus
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 02.12.2010; Public Library of Science (PLoS)
• Subjects: Acids; Amino acids; Antigens, CD - metabolism; Biology; CD44 antigen; CD81 antigen; Cell adhesion & migration; Cell culture; Cell Membrane - metabolism; Dendritic cells; Endoplasmic reticulum; Endosomes - metabolism; Enzymes; Fibroblasts; Fibroblasts - metabolism; Gene therapy; HeLa Cells; Homology; Human papillomavirus; Humans; Hyaluronan Receptors - metabolism; Hyaluronic acid; Hyaluronic Acid - chemistry; Inflammation; Inflammatory diseases; Intracellular; Isotopes; Laboratories; Ligases; Lysosomes; Lysosomes - metabolism; Membrane Glycoproteins - metabolism; Membrane Proteins - metabolism; Models, Biological; Neutrophils; Physiology; Protein turnover; Proteins; Proteome; Proteomics; Proteomics - methods; Stable isotopes; Tandem Mass Spectrometry - methods; Tetraspanin 28; Tetraspanin-29; Transmembrane domains; Trends; Ubiquitin; Ubiquitin-protein ligase; United States > US; Maryland
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0015132

Membrane-associated RING-CH (MARCH) proteins represent a family of transmembrane ubiquitin ligases modulating intracellular trafficking and turnover of transmembrane protein targets. While homologous proteins encoded by gamma-2 herpesviruses and leporipoxviruses have been studied extensively, limited information is available regarding the physiological targets of cellular MARCH proteins. To identify host cell proteins targeted by the human MARCH-VIII ubiquitin ligase we used stable isotope labeling of amino-acids in cell culture (SILAC) to monitor MARCH-dependent changes in the membrane proteomes of human fibroblasts. Unexpectedly, we observed that MARCH-VIII reduced the surface expression of Bap31, a chaperone that predominantly resides in the endoplasmic reticulum (ER). We demonstrate that Bap31 associates with the transmembrane domains of several MARCH proteins and controls intracellular transport of MARCH proteins. In addition, we observed that MARCH-VIII reduced the surface expression of the hyaluronic acid-receptor CD44 and both MARCH-VIII and MARCH-IV sequestered the tetraspanin CD81 in endo-lysosomal vesicles. Moreover, gene knockdown of MARCH-IV increased surface levels of endogenous CD81 suggesting a constitutive involvement of this family of ubiquitin ligases in the turnover of tetraspanins. Our data thus suggest a role of MARCH-VIII and MARCH-IV in the regulated turnover of CD81 and CD44, two ubiquitously expressed, multifunctional proteins.