Antisense promoter mapping. Inhibitory methods of transcriptional analysis.
We have employed antisense methods to study the transcriptional functions of c-fos protein (Fos). Clones expressing inducible anti-fos RNA have been employed to inhibit c-fos expression, resulting in activation of c-fos transcription by inhibiting its normal repressor function. The sites of negative regulation by Fos have been mapped using this antisense mapping method which demonstrates that the serum response element represents the major site of repression by endogenous c-fos protein. A similar strategy (antisense cloning) has been employed to clone four target genes that are Fos dependent. These cDNAs encode mRNAs that are rapidly induced by serum (although this induction is blocked by cycloheximide) but are blocked by induction of anti-fos RNA. These inhibitory methods of studying transcription factor function are extremely useful for transcription factors (like Fos) that require cooperation with other factors to modulate gene transcription.
Holt
J T
JT
Department of Cell Biology, Vanderbilt University, Nashville, Tennessee 37232.
D016375:Antisense Elements (Genetics); D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular; D016762:Genes, fos / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016372:RNA, Antisense / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic / Q000187:drug effects*
D016375:Antisense Elements (Genetics); D016372:RNA, Antisense; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase