Dissecting the activating mutations in v-erbB of avian erythroblastosis virus strain R.
The v-erbB oncogene isolated from the R (or ES4) strain of avian erythroblastosis virus is capable of inducing erythroleukemia and fibrosarcomas. This oncogene differs from the proto-oncogene c-erbB, the avian homolog of the epidermal growth factor receptor, by its lack of an intact ligand-binding domain as well as additional alterations in its cytoplasmic coding sequences. By contrast, the insertionally activated c-erbB, a variant oncogene, which encodes a product that also lacks the ligand-binding domain but is otherwise unaltered in its cytoplasmic coding sequences, is capable of inducing leukemia but cannot induce sarcomas. In this report, we show that the critical changes for activating the sarcomagenic potential displayed by v-erbB R are two point mutations within the tyrosine kinase domain and an internal deletion of 21 amino acids in the carboxyl-terminal regulatory domain. The removal of the carboxyl-terminal autophosphorylation sites is not obligatory. These activating mutations (Arg-263 to His, Ile-384 to Ser, and the deletion of residues 494 to 514), when introduced singly into the insertionally activated c-erbB, all dramatically increase fibroblast-transforming potential. Arg-263 resides near the highly conserved HRD motif of the kinase domain, and its mutation to His increases the autophosphorylation activity. The other two mutations do not alter the intrinsic kinase activity and presumably affect other aspects of the receptor involved in growth signaling. Therefore, the high transforming potential of v-erbB R is a consequence of synergism among multiple activating mutations.
Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio 44106.
H K
HK
Shu
R J
RJ
Pelley
H J
HJ
Kung
D001355:Alpharetrovirus / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002642:Chick Embryo; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D018739:Oncogene Proteins v-erbB; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D009857:Oncogenes; D010957:Plasmids; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D011994:Recombinant Proteins / Q000302:isolation & purification; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012513:Sarcoma, Experimental / Q000235:genetics / Q000382:microbiology*; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
C000715470:MAS1 protein, human; D009838:Oligodeoxyribonucleotides; D018739:Oncogene Proteins v-erbB; D009856:Oncogene Proteins, Viral; D000090063:Proto-Oncogene Mas; D011994:Recombinant Proteins; D015682:Retroviridae Proteins, Oncogenic; D011505:Protein-Tyrosine Kinases