Interactions of Drosophila Cbl with epidermal growth factor receptors and role of Cbl in R7 photoreceptor cell development
dc.contributor.author | Meisner, Herman | |
dc.contributor.author | Daga, Andrea | |
dc.contributor.author | Buxton, Joanne M. | |
dc.contributor.author | Fernandez, Belen | |
dc.contributor.author | Chawla, Anil | |
dc.contributor.author | Banerjee, Utpal | |
dc.contributor.author | Czech, Michael P. | |
dc.date | 2022-08-11T08:09:33.000 | |
dc.date.accessioned | 2022-08-23T16:35:33Z | |
dc.date.available | 2022-08-23T16:35:33Z | |
dc.date.issued | 1997-04-01 | |
dc.date.submitted | 2009-03-24 | |
dc.identifier.citation | Mol Cell Biol. 1997 Apr;17(4):2217-25. | |
dc.identifier.issn | 0270-7306 (Print) | |
dc.identifier.pmid | 9121472 | |
dc.identifier.uri | http://hdl.handle.net/20.500.14038/38587 | |
dc.description.abstract | The human proto-oncogene product c-Cbl and a similar protein in Caenorhabditis elegans (Sli-1) contain a proline-rich COOH-terminal region that binds Src homology 3 (SH3) domains of proteins such as the adapter Grb2. Cb1-Grb2 complexes can be recruited to tyrosine-phosphorylated epidermal growth factor (EGF) receptors through the SH2 domain of Grb2. Here we identify by molecular cloning a Drosophila cDNA encoding a protein (Drosophila Cbl [D-Cbl]) that shows high sequence similarity to the N-terminal region of human c-Cbl but lacks proline-rich sequences and fails to bind Grb2. Nonetheless, in COS-1 cells, expression of hemagglutinin epitope-tagged D-Cbl results in its coimmunoprecipitation with EGF receptors in response to EGF. EGF also caused tyrosine phosphorylation of D-Cbl in such cells, but no association of phosphatidylinositol 3-kinase was detected in assays using anti-p85 antibody. A point mutation in D-Cbl (G305E) that suppresses the negative regulation of LET-23 by the Cbl homolog Sli-1 in C. elegans prevented tyrosine phosphorylation of D-Cbl as well as binding to the liganded EGF receptor in COS-1 cells. Colocalization of EGF receptors with both endogenous c-Cbl or expressed D-Cbl in endosomes of EGF-treated COS-1 cells is also demonstrated by immunofluorescence microscopy. In lysates of adult transgenic Drosophila melanogaster, GST-DCbl binds to the tyrosine-phosphorylated 150-kDa torso-DER chimeric receptor. Expression of D-Cbl directed by the sevenless enhancer in intact Drosophila compromises severely the development of the R7 photoreceptor neuron. These data suggest that despite the lack of Grb2 binding sites, D-Cbl functions as a negative regulator of receptor tyrosine kinase signaling in the Drosophila eye by a mechanism that involves its association with EGF receptors or other tyrosine kinases. | |
dc.language.iso | en_US | |
dc.relation | <a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=9121472&dopt=Abstract">Link to Article in PubMed</a> | |
dc.subject | *Adaptor Proteins, Signal Transducing | |
dc.subject | Amino Acid Sequence | |
dc.subject | Animals | |
dc.subject | Base Sequence | |
dc.subject | Binding Sites | |
dc.subject | Caenorhabditis elegans | |
dc.subject | DNA Primers | |
dc.subject | DNA, Complementary | |
dc.subject | Drosophila melanogaster | |
dc.subject | GRB2 Adaptor Protein | |
dc.subject | Humans | |
dc.subject | Molecular Sequence Data | |
dc.subject | Photoreceptors, Invertebrate | |
dc.subject | Protein Binding | |
dc.subject | Proteins | |
dc.subject | Proto-Oncogene Proteins | |
dc.subject | Proto-Oncogene Proteins c-cbl | |
dc.subject | Proto-Oncogenes | |
dc.subject | Receptor, Epidermal Growth Factor | |
dc.subject | Sequence Homology, Amino Acid | |
dc.subject | *Ubiquitin-Protein Ligases | |
dc.subject | src Homology Domains | |
dc.subject | Life Sciences | |
dc.subject | Medicine and Health Sciences | |
dc.title | Interactions of Drosophila Cbl with epidermal growth factor receptors and role of Cbl in R7 photoreceptor cell development | |
dc.type | Journal Article | |
dc.source.journaltitle | Molecular and cellular biology | |
dc.source.volume | 17 | |
dc.source.issue | 4 | |
dc.identifier.legacyfulltext | https://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2451&context=oapubs&unstamped=1 | |
dc.identifier.legacycoverpage | https://escholarship.umassmed.edu/oapubs/1452 | |
dc.identifier.contextkey | 794949 | |
refterms.dateFOA | 2022-08-23T16:35:34Z | |
html.description.abstract | <p>The human proto-oncogene product c-Cbl and a similar protein in Caenorhabditis elegans (Sli-1) contain a proline-rich COOH-terminal region that binds Src homology 3 (SH3) domains of proteins such as the adapter Grb2. Cb1-Grb2 complexes can be recruited to tyrosine-phosphorylated epidermal growth factor (EGF) receptors through the SH2 domain of Grb2. Here we identify by molecular cloning a Drosophila cDNA encoding a protein (Drosophila Cbl [D-Cbl]) that shows high sequence similarity to the N-terminal region of human c-Cbl but lacks proline-rich sequences and fails to bind Grb2. Nonetheless, in COS-1 cells, expression of hemagglutinin epitope-tagged D-Cbl results in its coimmunoprecipitation with EGF receptors in response to EGF. EGF also caused tyrosine phosphorylation of D-Cbl in such cells, but no association of phosphatidylinositol 3-kinase was detected in assays using anti-p85 antibody. A point mutation in D-Cbl (G305E) that suppresses the negative regulation of LET-23 by the Cbl homolog Sli-1 in C. elegans prevented tyrosine phosphorylation of D-Cbl as well as binding to the liganded EGF receptor in COS-1 cells. Colocalization of EGF receptors with both endogenous c-Cbl or expressed D-Cbl in endosomes of EGF-treated COS-1 cells is also demonstrated by immunofluorescence microscopy. In lysates of adult transgenic Drosophila melanogaster, GST-DCbl binds to the tyrosine-phosphorylated 150-kDa torso-DER chimeric receptor. Expression of D-Cbl directed by the sevenless enhancer in intact Drosophila compromises severely the development of the R7 photoreceptor neuron. These data suggest that despite the lack of Grb2 binding sites, D-Cbl functions as a negative regulator of receptor tyrosine kinase signaling in the Drosophila eye by a mechanism that involves its association with EGF receptors or other tyrosine kinases.</p> | |
dc.identifier.submissionpath | oapubs/1452 | |
dc.contributor.department | Program in Molecular Medicine | |
dc.source.pages | 2217-25 |