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Identification and characterization of a novel cytoskeleton-associated pp60src substrate
H Wu1, A B Reynolds, S B Kanner
1Department of Microbiology, Health Sciences Center, Charlottesville, Virginia 22908.
Abstract:
Transformation of cells by the src oncogene results in elevated tyrosine phosphorylation of two related proteins, p80 and p85 (p80/85). Immunostaining with specific monoclonal antibodies revealed a striking change of subcellular localization of p80/85 in src-transformed cells. p80/85 colocalizes with F-actin in peripheral extensions of normal cells and rosettes (podosomes) of src-transformed cells. Sequence analysis of cDNA clones encoding p80/85 revealed an amino-terminal domain composed of six copies of a direct tandem repeat, each repeat containing 37 amino acids, a carboxyl-terminal SH3 domain, and an interdomain region composed of a highly charged acidic region and a region rich in proline, serine, and threonine. The multidomain structure of p80/85 and its colocalization with F-actin in normal and src-transformed cells suggest that these proteins may associate with components of the cytoskeleton and contribute to organization of cell structure.
Insights
Src oncogene transformation alters the location of p80/85 proteins in cells. These proteins associate with F-actin, suggesting a role in cell structure organization.
Area of Science:
- Cell Biology
- Molecular Biology
- Oncogenesis
Background:
- The src oncogene is implicated in cellular transformation.
- Tyrosine phosphorylation of specific proteins is altered during transformation.
- The subcellular localization of proteins can change upon oncogenic transformation.
Purpose of the Study:
- To investigate the changes in p80/85 protein localization during src-mediated cellular transformation.
- To elucidate the structural features and potential functions of p80/85 proteins.
Main Methods:
- Immunostaining with monoclonal antibodies to detect p80/85 protein.
- Subcellular localization analysis using microscopy.
- Sequence analysis of cDNA clones encoding p80/85 proteins.
Main Results:
- Src oncogene transformation increases tyrosine phosphorylation of p80/85 proteins.
- p80/85 proteins exhibit altered subcellular localization, associating with F-actin in transformed cells.
- p80/85 proteins possess a multidomain structure including tandem repeats and an SH3 domain.
Conclusions:
- p80/85 proteins are dynamically regulated during src-mediated transformation.
- The association of p80/85 with F-actin suggests a role in cytoskeletal organization.
- These findings provide insights into the molecular mechanisms of oncogenic transformation and cell structure regulation.
