A host-dependent temperature-sensitive mutant of Rous sarcoma virus: evidence for host factors affecting transformation.
We have characterized a host range mutant of Rous sarcoma virus in order to identify host cell factors involved in transformation. This mutant, tsLA33-1, which was isolated from a stock of the temperature-sensitive mutant tsLA33, is not temperature-sensitive for transformation of chicken embryo fibroblasts, as judged by its ability to induce morphological changes and agar colony formation at both 36 and 41.5 degrees. In Rat-3 cells, however, this mutant induced a temperature-dependent transformation: infected Rat-3 cells were transformed at 34 degrees but not at 39.5 degrees. Retransformants were isolated from tsLA33-1-infected Rat-3 cells by growth in agar suspension at 39.5 degrees. Virus rescued from these retransformants induced a temperature-dependent transformation when reintroduced into rat cells. The level of expression of pp60v-src at 39.5 degrees was unchanged in the retransformants. When the retransformants were treated with herbimycin, an antibiotic which induces turnover of certain protein-tyrosine kinases, they reverted to a normal phenotype, indicating that the transformed phenotype of the retransformants was dependent on continued expression of pp60v-src. The retransformants are therefore pseudorevertants in which a cellular alteration has occurred that allows transformation at 39.5 degrees by the mutant pp60v-src. Thus the temperature-dependence of transformation by tsLA33-1 is affected by the cellular environment, and is suppressed or complemented both in chicken cells and in the rat cell pseudorevertants. No clear correlation between levels of phosphorylation at tyrosine and transformation was observed. In Rat-3 cells the pp60v-src encoded by tsLA33-1 may be defective in its interaction with low abundance substrates that are critical for transformation; alternatively the nonpermissive cells may require a higher threshold dose of pp60v-src for transformation.
Young
J C
JC
Department of Zoology, University of California, Berkeley 94720.
Liebl
E
E
Martin
G S
GS
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000254:growth & development*; D016227:Benzoquinones; D002460:Cell Line; D002472:Cell Transformation, Viral; D002642:Chick Embryo; D047029:Lactams, Macrocyclic; D009154:Mutation; D015688:Oncogene Protein pp60(v-src); D009856:Oncogene Proteins, Viral / Q000502:physiology; D009857:Oncogenes; D010750:Phosphoproteins / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D011809:Quinones / Q000494:pharmacology; D012367:RNA, Viral / Q000235:genetics; D051381:Rats; D012191:Retroviridae Proteins / Q000502:physiology; D017828:Rifabutin / Q000031:analogs & derivatives; D013696:Temperature
D016227:Benzoquinones; D047029:Lactams, Macrocyclic; D009856:Oncogene Proteins, Viral; D010750:Phosphoproteins; D011809:Quinones; D012367:RNA, Viral; D012191:Retroviridae Proteins; D017828:Rifabutin; C020754:herbimycin; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src)