A recombinant murine retrovirus vector containing the v-erbB gene of avian erythroblastosis virus was constructed to investigate v-erbB as a transforming gene for mammalian cells. A restriction fragment containing the v-erbB sequences from a molecular clone of avian erythroblastosis virus was inserted into a Moloney murine leukemia virus vector. The construct, designated MuLV/erbB, transformed NIH 3T3 cells at a high efficiency in the DNA transfection assay. Individual MuLV/erbB transfectants grew in soft agar and were tumorigenic. The transfectants contained v-erbB DNA sequences, expressed v-erbB-specific transcripts, and synthesized v-erbB-related glycoproteins. The majority of transfectants produced two major v-erbB gene products of 58 and 66 kilodaltons. However, some transfectants produced much smaller v-erbB-specific proteins. Tunicamycin experiments revealed that the size heterogeneity observed between different transfectants was not due to variations in glycoprotein processing, implying that, in some cases, alterations in the MuLV/erbB genome occurred during the transfection process. These findings indicate that expression of the complete v-erbB gene product is not required for transformation of NIH 3T3 cells. A transmissible murine v-erbB (M-erbB) virus was generated by infection of nonproducer transfectants with amphotrophic murine leukemia virus. Transmission of the rescued M-erbB virus was confirmed by DNA, RNA, and protein analyses. The introduction of a transforming v-erbB gene into mammalian cells by virus infection provides a means of analyzing the mechanism by which this epidermal growth factor receptor-related gene alters the growth and differentiation of cells from various lineages.
|Titolo:||Mammalian cell transformation by a murine retrovirus vector containing the avian erythroblastosis virus erbB gene|
|Parole Chiave:||Alpharetrovirus; Animals; Viral Proteins; Moloney murine leukemia virus; Mink; Humans; Mice; Rats; Avian leukosis virus; Oncogene Proteins, Viral; Transfection; Genetic Vectors; Species Specificity; Cell Transformation, Viral; Cell Line|
|Settore Scientifico Disciplinare:||Settore MED/04 - Patologia Generale|
|Data di pubblicazione:||ott-1986|
|Appare nelle tipologie:||01 - Articolo su periodico|