Functional analysis of the mouse H-2Kb gene promoter in embryonal carcinoma cells.
Mouse embryonal carcinoma (EC) cells do not express the major H-2 class I transplantation antigens. The latter, however, become detectable upon in vitro differentiation of EC cells. Neither class I H-2 genes nor the gene coding for beta-2 microglobulin (beta 2m) is transcribed in EC cells. We have constructed two hybrid plasmids containing the 5' flanking region of an H-2Kb gene followed by the coding regions of either the herpes simplex virus thymidine kinase (H-2 tk) or the chloramphenicol acetyl transferase (H-2 CAT) genes. Upon transfer into EC cells, the H-2 tk hybrid gene is expressed in F9 tk- cell lines which thus acquire a stable tk+ phenotype. When such transformed clones are induced to differentiate in vitro, tk activity shows a moderate increase, which reflects an increase in transcription of the hybrid gene. In transient transformation experiments, EC cells were found to express the H-2 CAT hybrid gene as well. We conclude that the 2 kilobase pair region of the H-2Kb gene which we used contains an active promoter region, but does not include all the elements required for the correct regulation of the H-2Kb gene.
Daniel-Vedele
F
F
Israel
A
A
Benicourt
C
C
Kourilsky
P
P
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase; D005796:Genes; D005802:Genes, MHC Class II; D005814:Genes, Viral; D006183:H-2 Antigens / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013724:Teratoma / Q000276:immunology*; D013937:Thymidine Kinase / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D001613:beta 2-Microglobulin / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D001613:beta 2-Microglobulin; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; D013937:Thymidine Kinase; D001616:beta-Galactosidase