Vaccinia virus systems for expression of G alpha genes in S49 cells.
In this chapter, we describe the use of recombinant VV to express G alpha subunits efficiently in S49 cyc- cells, a murine lymphoma cell line deficient in endogenous Gs alpha activity. Additional studies have shown that VV infection does not interfere with G-protein-coupled signal transduction events. In particular, the VV-expressed rat Gs alpha subunit is able to mediate efficient receptor-dependent and receptor-independent activation of adenylyl cyclase in cyc- cells. This system should therefore be useful for rapidly assessing the activity of modified or chimeric Gs alpha subunits.
Quan
F
F
Vollum Institute, Oregon Health Sciences University, Portland 97201.
Forte
M
M
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D004330:Drosophila; D019204:GTP-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008223:Lymphoma; D008322:Mammals; D051379:Mice; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*; D014616:Vaccinia virus
D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; C020888:1,2-dielaidoylphosphatidylethanolamine; D019204:GTP-Binding Proteins