Antisense mapping of the c-fos promoter: role of the serum response element.
Using an antisense RNA approach to eliminate endogenous expression of the c-fos protein, we have verified by nuclear run-on and transient expression assays that the Fos protein is a negative regulator of its own transcription in vivo. The negative autoregulation of the c-fos promoter by Fos was further confirmed by overexpression of an antisense-resistant c-fos expressing vector. Antisense mapping of the c-fos promoter demonstrated that the serum responsive element (SRE) represents the major site for c-fos suppression only during the first hour, but that additional adjacent DNA sequences are required for suppression at later times. We propose that antisense inhibition of transcriptional repressors provides a useful method for analyzing the significance and mechanism of transcriptional repression in vivo.
Robinson-Benion
C
C
Department of Cell Biology, Vanderbilt University School of Medicine, Nashville, TN 37232.
Kamata
N
N
Holt
J T
JT
D016475:3T3 Cells; D000818:Animals; D002467:Cell Nucleus / Q000502:physiology; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D002874:Chromosome Mapping / Q000379:methods; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D016762:Genes, fos / Q000235:genetics*; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016760:Proto-Oncogene Proteins c-fos / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D016372:RNA, Antisense / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D012097:Repressor Proteins / Q000235:genetics / Q000502:physiology; D026362:Serum Response Factor; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection / Q000235:genetics
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016760:Proto-Oncogene Proteins c-fos; D016372:RNA, Antisense; D012097:Repressor Proteins; D026362:Serum Response Factor; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase