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dc.contributor.authorChakrabarti, Ranjan
dc.contributor.authorJoly, Marguerite
dc.contributor.authorCorvera, Silvia
dc.date2022-08-11T08:10:05.000
dc.date.accessioned2022-08-23T16:55:00Z
dc.date.available2022-08-23T16:55:00Z
dc.date.issued1993-10-01
dc.date.submitted2008-10-31
dc.identifier.citationJ Cell Biol. 1993 Oct;123(1):79-87.
dc.identifier.issn0021-9525 (Print)
dc.identifier.pmid8408208
dc.identifier.urihttp://hdl.handle.net/20.500.14038/42625
dc.description.abstractMechanisms for intracellular retention of proteins are induced during adipocytic differentiation of 3T3-L1 cells. To investigate the potential role of clathrin lattices in these retention processes, we performed a morphological and biochemical analysis of coated vesicle components in 3T3-L1 cells. Optical sectioning and image restoration revealed a marked increase in the staining of clathrin and beta adaptins in the perinuclear region of cells with differentiation. In addition, predominance of beta (subunit of the AP-2, plasma membrane adaptor) over beta' (subunit of the AP-1, Golgi adaptor) adaptin was observed in immunoblots of clathrin-coated vesicles purified from nondifferentiated fibroblasts, and this ratio was reversed in coated vesicles purified from differentiated adipocytes. These results indicate that the relative abundance of TGN-derived clathrin lattices increases markedly during adipocytic differentiation. Subcellular fractionation indicated that cytosolic AP-1 and AP-2 adaptors comprised approximately 70% of the total cellular adaptor pool. Interestingly, neither the concentration nor the relative ratio of cytosolic AP-1 to AP-2 adaptors increased significantly during differentiation. These data suggest that the increase in TGN-derived lattices results from differentiation-induced mechanisms for enhanced assembly or stabilization of adaptors on Golgi membranes. Interestingly, double-immunofluorescence microscopy also revealed that whereas extensive colocalization between clathrin and beta adaptins occurred both in fibroblasts and adipocytes, structures stained only with anti-adaptin antibody could be detected. Taken together these results suggest that membranes coated with adaptors, but not clathrin, can exist in these cells.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=8408208&dopt=Abstract">Link to Article in PubMed</a>
dc.subject3T3 Cells
dc.subjectAdaptor Protein Complex alpha Subunits
dc.subjectAdaptor Protein Complex gamma Subunits
dc.subjectAdaptor Proteins, Vesicular Transport
dc.subjectAdipocytes
dc.subjectAnimals
dc.subjectCell Compartmentation
dc.subject*Cell Differentiation
dc.subject*Clathrin
dc.subjectCoated Pits, Cell-Membrane
dc.subjectCytosol
dc.subjectFibroblasts
dc.subjectFluorescent Antibody Technique
dc.subjectGolgi Apparatus
dc.subjectIntracellular Membranes
dc.subjectMice
dc.subjectProteins
dc.subjectSubcellular Fractions
dc.subjectCell Biology
dc.subjectMolecular Biology
dc.titleRedistribution of clathrin-coated vesicle adaptor complexes during adipocytic differentiation of 3T3-L1 cells
dc.typeJournal Article
dc.source.journaltitleThe Journal of cell biology
dc.source.volume123
dc.source.issue1
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1955&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/956
dc.identifier.contextkey659137
refterms.dateFOA2022-08-23T16:55:01Z
html.description.abstract<p>Mechanisms for intracellular retention of proteins are induced during adipocytic differentiation of 3T3-L1 cells. To investigate the potential role of clathrin lattices in these retention processes, we performed a morphological and biochemical analysis of coated vesicle components in 3T3-L1 cells. Optical sectioning and image restoration revealed a marked increase in the staining of clathrin and beta adaptins in the perinuclear region of cells with differentiation. In addition, predominance of beta (subunit of the AP-2, plasma membrane adaptor) over beta' (subunit of the AP-1, Golgi adaptor) adaptin was observed in immunoblots of clathrin-coated vesicles purified from nondifferentiated fibroblasts, and this ratio was reversed in coated vesicles purified from differentiated adipocytes. These results indicate that the relative abundance of TGN-derived clathrin lattices increases markedly during adipocytic differentiation. Subcellular fractionation indicated that cytosolic AP-1 and AP-2 adaptors comprised approximately 70% of the total cellular adaptor pool. Interestingly, neither the concentration nor the relative ratio of cytosolic AP-1 to AP-2 adaptors increased significantly during differentiation. These data suggest that the increase in TGN-derived lattices results from differentiation-induced mechanisms for enhanced assembly or stabilization of adaptors on Golgi membranes. Interestingly, double-immunofluorescence microscopy also revealed that whereas extensive colocalization between clathrin and beta adaptins occurred both in fibroblasts and adipocytes, structures stained only with anti-adaptin antibody could be detected. Taken together these results suggest that membranes coated with adaptors, but not clathrin, can exist in these cells.</p>
dc.identifier.submissionpathoapubs/956
dc.contributor.departmentDepartment of Cell Biology
dc.contributor.departmentProgram in Molecular Medicine
dc.source.pages79-87


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