A small v-sis/platelet-derived growth factor (PDGF) B-protein domain in which subtle conformational changes abrogate PDGF receptor interaction and transforming activity.
Deletion scanning mutagenesis within the transforming region of the v-sis oncogene was used to dissect structure-function relationships. Mutations affecting codons within a domain encoding amino acids 136 through 148 had no effect upon homodimer formation or recognition by antisera which detect determinants dependent upon native intrachain disulfide linkages, yet the same mutations completely abolished transforming activity. A platelet-derived growth factor B (PDGF B) monoclonal antibody that prevents its interaction with PDGF receptors recognized v-sis, delta 142 (deletion of codon 142), and delta 148 but not delta 136, delta 137, or delta 139 mutants. These findings mapped the epitope recognized by this monoclonal antibody to include amino acid residues 136 to 139. Furthermore, mutations in the codon 136 to 148 domain caused markedly impaired ability to induce PDGF receptor tyrosine phosphorylation. Thus, subtle conformational alterations in this small domain critically affect PDGF receptor recognition and/or functional activation.
Laboratory of Cellular and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
N
N
Giese
W J
WJ
LaRochelle
M
M
May-Siroff
K C
KC
Robbins
S A
SA
Aaronson
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002471:Cell Transformation, Neoplastic; D003062:Codon; D004252:DNA Mutational Analysis; D000939:Epitopes; D051379:Mice; D020575:Oncogene Proteins v-sis; D019000:Phosphotyrosine; D010982:Platelet-Derived Growth Factor / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000502:physiology*; D017479:Receptors, Platelet-Derived Growth Factor; D015682:Retroviridae Proteins, Oncogenic / Q000502:physiology*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D000911:Antibodies, Monoclonal; D003062:Codon; D000939:Epitopes; D020575:Oncogene Proteins v-sis; D010982:Platelet-Derived Growth Factor; D011956:Receptors, Cell Surface; D015682:Retroviridae Proteins, Oncogenic; D019000:Phosphotyrosine; D014443:Tyrosine; D017479:Receptors, Platelet-Derived Growth Factor