Demonstration in living cells of an intragenic negative regulatory element within the rodent c-fos gene.
We studied c-fos gene expression in rat fibroblasts by microinjection of regulatory DNA sequences, such as the serum response element (SRE) present in c-fos promotor, in order to compete directly with such sequences for binding of putative regulatory factors. We show that an additional fos intragenic regulatory element (FIRE) is located at the end of exon 1. When coinjected with an SRE oligonucleotide, it induced c-fos expression in quiescent cells, whereas injection of SRE sequence alone failed to do so. Moreover, injection in quiescent cells of an SRE oligonucleotide together with a p-fos-lacZ construct containing the c-fos SRE as well as an in-frame insertion of FIRE resulted in a block to beta-galactosidase expression that can be relieved by coinjection of the FIRE sequence.
Lamb
N J
NJ
Laboratoire de Biologie Moléculaire, URA CNRS 1191, Montpellier, France.
Fernandez
A
A
Tourkine
N
N
Jeanteur
P
P
Blanchard
J M
JM
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003850:Deoxyribonuclease I; D005091:Exons; D015870:Gene Expression; D005809:Genes, Regulator; D006801:Humans; D007438:Introns; D008845:Microinjections; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016760:Proto-Oncogene Proteins c-fos; D011519:Proto-Oncogenes; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects
D015345:Oligonucleotide Probes; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D011505:Protein-Tyrosine Kinases; D003850:Deoxyribonuclease I