Ubiquitylation of leptin receptor OB-Ra regulates its clathrin-mediated endocytosis
Leptin receptors are constitutively endocytosed in a ligand‐independent manner. To study their endocytosis, leptin receptors OB‐Ra and OB‐Rb were expressed in HeLa cells. Both receptor isoforms were ubiquitylated, internalized by clathrin‐mediated endocytosis and transported to Hrs‐positive endosome...
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| Veröffentlicht in: | The EMBO journal 2006-03, Vol.25 (5), p.932-942 |
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| description | Leptin receptors are constitutively endocytosed in a ligand‐independent manner. To study their endocytosis, leptin receptors OB‐Ra and OB‐Rb were expressed in HeLa cells. Both receptor isoforms were ubiquitylated, internalized by clathrin‐mediated endocytosis and transported to Hrs‐positive endosomes after their internalization. Proteasome inhibitors inhibited OB‐Ra but not OB‐Rb internalization from the cell surface. OB‐Ra ubiquitylation occurred on lysine residues K877 and K889 in the cytoplasmic tail, the mutation of which abolished OB‐Ra internalization. Fusion of an ubiquitin molecule at the C‐terminus of an OB‐Ra construct defective both in ubiquitylation and endocytosis restored clathrin‐dependent endocytosis of the receptor. The internalization of this constitutively mono‐ubiquitylated construct was no longer sensitive to proteasome inhibitors, which inhibited OB‐Ra endocytosis by blocking its ubiquitylation. Fusion of an ubiquitin molecule to a transferrin receptor deleted from its own endocytosis motif restored clathrin‐mediated endocytosis. We propose that mono‐ubiquitin conjugates act as internalization motifs for clathrin‐dependent endocytosis of leptin receptor OB‐Ra. |
| doi_str_mv | 10.1038/sj.emboj.7600989 |
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To study their endocytosis, leptin receptors OB‐Ra and OB‐Rb were expressed in HeLa cells. Both receptor isoforms were ubiquitylated, internalized by clathrin‐mediated endocytosis and transported to Hrs‐positive endosomes after their internalization. Proteasome inhibitors inhibited OB‐Ra but not OB‐Rb internalization from the cell surface. OB‐Ra ubiquitylation occurred on lysine residues K877 and K889 in the cytoplasmic tail, the mutation of which abolished OB‐Ra internalization. Fusion of an ubiquitin molecule at the C‐terminus of an OB‐Ra construct defective both in ubiquitylation and endocytosis restored clathrin‐dependent endocytosis of the receptor. The internalization of this constitutively mono‐ubiquitylated construct was no longer sensitive to proteasome inhibitors, which inhibited OB‐Ra endocytosis by blocking its ubiquitylation. Fusion of an ubiquitin molecule to a transferrin receptor deleted from its own endocytosis motif restored clathrin‐mediated endocytosis. We propose that mono‐ubiquitin conjugates act as internalization motifs for clathrin‐dependent endocytosis of leptin receptor OB‐Ra.</description><identifier>ISSN: 0261-4189</identifier><identifier>EISSN: 1460-2075</identifier><identifier>DOI: 10.1038/sj.emboj.7600989</identifier><identifier>PMID: 16482222</identifier><identifier>CODEN: EMJODG</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Cell Membrane ; Clathrin - metabolism ; EMBO20 ; Endocytosis ; endocytosis motif ; Endosomes - metabolism ; Enzyme Inhibitors - pharmacology ; Fusion ; Gene expression ; HeLa Cells ; Humans ; Inhibitors ; leptin receptor ; Mutation ; Mutation - genetics ; Neurons ; proteasome ; Proteasome Inhibitors ; Protein Isoforms ; Receptors, Cell Surface - genetics ; Receptors, Cell Surface - metabolism ; Receptors, Leptin ; Receptors, Transferrin - metabolism ; siRNA ; ubiquitin ; Ubiquitin - metabolism</subject><ispartof>The EMBO journal, 2006-03, Vol.25 (5), p.932-942</ispartof><rights>European Molecular Biology Organization 2006</rights><rights>Copyright © 2006 European Molecular Biology Organization</rights><rights>Copyright Nature Publishing Group Mar 8, 2006</rights><rights>Copyright © 2006, European Molecular Biology Organization 2006</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6399-cc49851dfb05fd46118071e91e9ffb19294d96f3d5e5497349d7a6aa3c0c541c3</citedby><cites>FETCH-LOGICAL-c6399-cc49851dfb05fd46118071e91e9ffb19294d96f3d5e5497349d7a6aa3c0c541c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1409713/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1409713/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,1417,1433,27924,27925,41120,42189,45574,45575,46409,46833,51576,53791,53793</link.rule.ids><linktorsrc>$$Uhttps://doi.org/10.1038/sj.emboj.7600989$$EView_record_in_Springer_Nature$$FView_record_in_$$GSpringer_Nature</linktorsrc><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16482222$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Belouzard, Sandrine</creatorcontrib><creatorcontrib>Rouillé, Yves</creatorcontrib><title>Ubiquitylation of leptin receptor OB-Ra regulates its clathrin-mediated endocytosis</title><title>The EMBO journal</title><addtitle>EMBO J</addtitle><addtitle>EMBO J</addtitle><description>Leptin receptors are constitutively endocytosed in a ligand‐independent manner. To study their endocytosis, leptin receptors OB‐Ra and OB‐Rb were expressed in HeLa cells. Both receptor isoforms were ubiquitylated, internalized by clathrin‐mediated endocytosis and transported to Hrs‐positive endosomes after their internalization. Proteasome inhibitors inhibited OB‐Ra but not OB‐Rb internalization from the cell surface. OB‐Ra ubiquitylation occurred on lysine residues K877 and K889 in the cytoplasmic tail, the mutation of which abolished OB‐Ra internalization. Fusion of an ubiquitin molecule at the C‐terminus of an OB‐Ra construct defective both in ubiquitylation and endocytosis restored clathrin‐dependent endocytosis of the receptor. The internalization of this constitutively mono‐ubiquitylated construct was no longer sensitive to proteasome inhibitors, which inhibited OB‐Ra endocytosis by blocking its ubiquitylation. Fusion of an ubiquitin molecule to a transferrin receptor deleted from its own endocytosis motif restored clathrin‐mediated endocytosis. We propose that mono‐ubiquitin conjugates act as internalization motifs for clathrin‐dependent endocytosis of leptin receptor OB‐Ra.</description><subject>Cell Membrane</subject><subject>Clathrin - metabolism</subject><subject>EMBO20</subject><subject>Endocytosis</subject><subject>endocytosis motif</subject><subject>Endosomes - metabolism</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Fusion</subject><subject>Gene expression</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Inhibitors</subject><subject>leptin receptor</subject><subject>Mutation</subject><subject>Mutation - genetics</subject><subject>Neurons</subject><subject>proteasome</subject><subject>Proteasome Inhibitors</subject><subject>Protein Isoforms</subject><subject>Receptors, Cell Surface - genetics</subject><subject>Receptors, Cell Surface - metabolism</subject><subject>Receptors, Leptin</subject><subject>Receptors, Transferrin - metabolism</subject><subject>siRNA</subject><subject>ubiquitin</subject><subject>Ubiquitin - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The EMBO journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Belouzard, Sandrine</au><au>Rouillé, Yves</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ubiquitylation of leptin receptor OB-Ra regulates its clathrin-mediated endocytosis</atitle><jtitle>The EMBO journal</jtitle><stitle>EMBO J</stitle><addtitle>EMBO J</addtitle><date>2006-03-08</date><risdate>2006</risdate><volume>25</volume><issue>5</issue><spage>932</spage><epage>942</epage><pages>932-942</pages><issn>0261-4189</issn><eissn>1460-2075</eissn><coden>EMJODG</coden><abstract>Leptin receptors are constitutively endocytosed in a ligand‐independent manner. To study their endocytosis, leptin receptors OB‐Ra and OB‐Rb were expressed in HeLa cells. Both receptor isoforms were ubiquitylated, internalized by clathrin‐mediated endocytosis and transported to Hrs‐positive endosomes after their internalization. Proteasome inhibitors inhibited OB‐Ra but not OB‐Rb internalization from the cell surface. OB‐Ra ubiquitylation occurred on lysine residues K877 and K889 in the cytoplasmic tail, the mutation of which abolished OB‐Ra internalization. Fusion of an ubiquitin molecule at the C‐terminus of an OB‐Ra construct defective both in ubiquitylation and endocytosis restored clathrin‐dependent endocytosis of the receptor. The internalization of this constitutively mono‐ubiquitylated construct was no longer sensitive to proteasome inhibitors, which inhibited OB‐Ra endocytosis by blocking its ubiquitylation. Fusion of an ubiquitin molecule to a transferrin receptor deleted from its own endocytosis motif restored clathrin‐mediated endocytosis. We propose that mono‐ubiquitin conjugates act as internalization motifs for clathrin‐dependent endocytosis of leptin receptor OB‐Ra.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>16482222</pmid><doi>10.1038/sj.emboj.7600989</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Cell Membrane Clathrin - metabolism EMBO20 Endocytosis endocytosis motif Endosomes - metabolism Enzyme Inhibitors - pharmacology Fusion Gene expression HeLa Cells Humans Inhibitors leptin receptor Mutation Mutation - genetics Neurons proteasome Proteasome Inhibitors Protein Isoforms Receptors, Cell Surface - genetics Receptors, Cell Surface - metabolism Receptors, Leptin Receptors, Transferrin - metabolism siRNA ubiquitin Ubiquitin - metabolism |
| title | Ubiquitylation of leptin receptor OB-Ra regulates its clathrin-mediated endocytosis |
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