Structure and regulation of the growth arrest-specific (gas-1) promoter.
We have isolated recombinant genomic clones encompassing several kilobase pairs of the 5'-flanking regions of both the human and murine gas-1 gene (growth arrest-specific gene 1). Both species share a highly conserved region of approximately 550 base pairs upstream of the gas-1 transcription start site. Deletion analysis of the murine gas-1 promoter demonstrated that a fragment containing the first 665 base pairs is sufficient to drive the serum-regulated expression of a luciferase reporter gene in NIH3T3 cells, in a manner qualitatively reflecting the activity of the endogenous gene. Gel retardation assays indicated the presence of a number of DNA-binding proteins specific for sequences contained within the gas-1 transcription regulatory region. Comparative studies with extracts prepared from growing and resting cells revealed several growth state-specific binding activities. One promoter fragment that bound prominent growth- and arrest-specific complexes was further analyzed by copper-phenanthroline footprinting. It was found that the same DNA element is a target both for growth- and for arrest-specific activities. The factors characterized in this study are the first candidates for transcriptional regulators mediating cell growth-specific repression and/or growth arrest-specific activation of gene expression.
de Martin
R
R
European Molecular Biology Laboratory, Heidelberg, Germany.
Cowled
P A
PA
Smith
S E
SE
Papavassiliou
A G
AG
Sorrentino
V
V
Philipson
L
L
Bohmann
D
D
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000235:genetics; D017124:Conserved Sequence; D015971:Gene Expression Regulation, Enzymologic; D015698:Genomic Library; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D016385:TATA Box; D014158:Transcription, Genetic; D014162:Transfection
D008156:Luciferases