Transient gene expression by SV40 promoter characterizes sequential differentiation of embryonal carcinoma F9 cells into primitive and visceral endoderm.
Transient expression of the chloramphenicol acetyltransferase (CAT) gene under the control of simian virus 40 (SV40)y Moloney murine leukemia virus, human T cell leukemia virus, and cytomegalovirus promoters was stimulated by the differentiation of F9 stem cells into primitive endoderm, but repressed again by further differentiation into visceral endoderm. Deletion mutants of the SV40 enhancer showed that a similar set of motifs is critical for CAT expression at all stages of F9 differentiation, but differentiation dependency was observed even in their absence. The stability of transient gene expression under the control of the SV40 promoter was markedly dependent on F9 differentiation. Appreciable expression was detected even in undifferentiated F9 cells immediately after gene transfection, was maximal at 12 h and declined rapidly thereafter. On the other hand, expression in primitive endoderm increased until 72 h. The decline was accelerated again in visceral endoderm. This shift was somewhat specific to the virus promoter since CAT expression in undifferentiated F9 cells under the control of the elongation factor 1 alpha promoter was more stable than for virus promoters tested. Thus, the change in stability of expression is important for differentiation-dependent virus promoter activity.
Taniguchi
Y
Y
Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Japan.
Miki
K
K
Kitagawa
Y
Y
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics*; D054278:Embryonal Carcinoma Stem Cells; D004707:Endoderm; D015967:Gene Expression Regulation, Viral; D051379:Mice; D008969:Molecular Sequence Data; D014411:Neoplastic Stem Cells; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics*; D014407:Tumor Cells, Cultured
D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D015500:Chloramphenicol O-Acetyltransferase