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dc.contributor.authorShisheva, Assia
dc.contributor.authorSudhof, Thomas C.
dc.contributor.authorCzech, Michael P.
dc.date2022-08-11T08:09:33.000
dc.date.accessioned2022-08-23T16:35:36Z
dc.date.available2022-08-23T16:35:36Z
dc.date.issued1994-05-01
dc.date.submitted2009-03-24
dc.identifier.citationMol Cell Biol. 1994 May;14(5):3459-68.
dc.identifier.issn0270-7306 (Print)
dc.identifier.pmid7513052
dc.identifier.urihttp://hdl.handle.net/20.500.14038/38596
dc.description.abstractCellular mechanisms for controlling membrane trafficking appear to involve small GTP-binding proteins such as the Rab proteins. Rab function is regulated by GDP dissociation inhibitor (GDI), which releases Rab proteins from membranes and inhibits GDP dissociation. Here we report the isolation of a full-length cDNA encoding a novel GDI isoform of 445 amino acids (GDI-2) with a deduced molecular weight of 50,649 from mouse skeletal muscle. Full-length and partial cDNA clones encoding a previously reported GDI protein (GDI-1) were also isolated from cDNA libraries prepared from rat brain and mouse skeletal muscle, respectively. The degree of deduced amino acid sequence identity between mouse GDI-2 and our mouse GDI-1 cDNA clone is 86%. Northern (RNA blot) analysis revealed that in human tissues, both GDI-1 and GDI-2 transcripts were abundant in brain, skeletal muscle, and pancreas but were weakly expressed in heart and liver. GDI-1 mRNA was expressed in kidney, whereas GDI-2 was almost absent, while in lung the relative amounts of these mRNA species were reversed. Specific antibodies against mouse GDI-1 and GDI-2 based on unique peptide sequences in the proteins were raised. Differentiation of 3T3-L1 fibroblasts into highly insulin-responsive adipocytes was accompanied by large increases in both mRNA and protein levels of GDI-1 and GDI-2. GDI-1 and GDI-2 expressed as glutathione S-transferase fusion proteins were both able to solubilize the membrane-bound forms of Rab4 and Rab5 in a GDP/GTP-dependent manner. Taken together, these data demonstrate that the protein products of at least two genes regulate the membrane dynamics of Rab proteins in mice.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=7513052&dopt=Abstract">Link to Article in PubMed</a>
dc.subjectAmino Acid Sequence
dc.subjectAnimals
dc.subjectBase Sequence
dc.subjectBlotting, Northern
dc.subjectBrain
dc.subjectCell Line
dc.subjectCloning, Molecular
dc.subjectDNA Primers
dc.subjectGTP-Binding Proteins
dc.subjectGlutathione Transferase
dc.subject*Guanine Nucleotide Dissociation Inhibitors
dc.subjectHumans
dc.subjectLiver
dc.subjectMolecular Sequence Data
dc.subjectMuscles
dc.subjectMyocardium
dc.subjectOrgan Specificity
dc.subjectPancreas
dc.subjectPolymerase Chain Reaction
dc.subjectRNA
dc.subjectRNA, Messenger
dc.subjectRats
dc.subjectRecombinant Fusion Proteins
dc.subjectSequence Homology, Amino Acid
dc.subjectSequence Homology, Nucleic Acid
dc.subjectSpecies Specificity
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleCloning, characterization, and expression of a novel GDP dissociation inhibitor isoform from skeletal muscle
dc.typeJournal Article
dc.source.journaltitleMolecular and cellular biology
dc.source.volume14
dc.source.issue5
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2459&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/1460
dc.identifier.contextkey794957
refterms.dateFOA2022-08-23T16:35:36Z
html.description.abstract<p>Cellular mechanisms for controlling membrane trafficking appear to involve small GTP-binding proteins such as the Rab proteins. Rab function is regulated by GDP dissociation inhibitor (GDI), which releases Rab proteins from membranes and inhibits GDP dissociation. Here we report the isolation of a full-length cDNA encoding a novel GDI isoform of 445 amino acids (GDI-2) with a deduced molecular weight of 50,649 from mouse skeletal muscle. Full-length and partial cDNA clones encoding a previously reported GDI protein (GDI-1) were also isolated from cDNA libraries prepared from rat brain and mouse skeletal muscle, respectively. The degree of deduced amino acid sequence identity between mouse GDI-2 and our mouse GDI-1 cDNA clone is 86%. Northern (RNA blot) analysis revealed that in human tissues, both GDI-1 and GDI-2 transcripts were abundant in brain, skeletal muscle, and pancreas but were weakly expressed in heart and liver. GDI-1 mRNA was expressed in kidney, whereas GDI-2 was almost absent, while in lung the relative amounts of these mRNA species were reversed. Specific antibodies against mouse GDI-1 and GDI-2 based on unique peptide sequences in the proteins were raised. Differentiation of 3T3-L1 fibroblasts into highly insulin-responsive adipocytes was accompanied by large increases in both mRNA and protein levels of GDI-1 and GDI-2. GDI-1 and GDI-2 expressed as glutathione S-transferase fusion proteins were both able to solubilize the membrane-bound forms of Rab4 and Rab5 in a GDP/GTP-dependent manner. Taken together, these data demonstrate that the protein products of at least two genes regulate the membrane dynamics of Rab proteins in mice.</p>
dc.identifier.submissionpathoapubs/1460
dc.contributor.departmentProgram in Molecular Medicine
dc.source.pages3459-68


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