Evidence for a protein-trafficking gene that rescues the defective glucocorticoid-regulated transport and Golgi retention of mouse mammary tumor virus glycoproteins in a rat hepatoma cell-sorting variant.
Glucocorticoids regulate the trafficking of cell surface mouse mammary tumor virus (MMTV) glycoproteins in the virus-infected rat hepatoma cell line M1.54. The CR4 rat hepatoma sorting variant, which is derived from M1.54 cells by immunoselection, is uniquely defective in the glucocorticoid-regulated transport of MMTV glycoproteins. Indirect immunofluorescence of fixed permeabilized cells and subcellular fractionation of isolated microsomes revealed that variant CR4 cells retain the MMTV glycoproteins in Golgi-like membranes after glucocorticoid treatment. The variant CR4 phenotype can be complemented by interspecies cell fusions with human HepG2 hepatoma cells and by DNA rescue with genomic fragments isolated from either human or rat hepatoma cells. Transfected wild-type genomic fragments rescue the sorting defect in CR4 at a frequency consistant with a single genetic locus, whereas homologous transfection with CR4 genomic DNA has no effect. Thus, complementation of a rat hepatoma cell-sorting variant supports the existence of a novel protein-trafficking activity encoded by the human or rat genomes that acts in trans in the Golgi to selectively mediate the sorting of cell surface MMTV glycoproteins in glucocorticoid-treated cells.
Bravo
D A
DA
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Goodman
L J
LJ
Firestone
G L
GL
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000235:genetics*; D001692:Biological Transport; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology; D004247:DNA / Q000378:metabolism; D005816:Genetic Complementation Test; D014644:Genetic Variation; D005938:Glucocorticoids / Q000494:pharmacology*; D006023:Glycoproteins / Q000235:genetics*; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008324:Mammary Tumor Virus, Mouse / Q000032:analysis; D010641:Phenotype; D051381:Rats; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000235:genetics*
D000957:Antigens, Viral, Tumor; D005938:Glucocorticoids; D006023:Glycoproteins; D014759:Viral Envelope Proteins; D004247:DNA