Sequence requirements for premature transcription arrest within the first intron of the mouse c-fos gene.
A strong block to the elongation of nascent RNA transcripts by RNA polymerase II occurs in the 5' part of the mammalian c-fos proto-oncogene. In addition to the control of initiation, this mechanism contributes to transcriptional regulation of the gene. In vitro transcription experiments using nuclear extracts and purified transcription templates allowed us to map a unique arrest site within the mouse first intron 385 nucleotides downstream from the promoter. This position is in keeping with that estimated from nuclear run-on assays performed with short DNA probes and thus suggests that it corresponds to the actual block in vivo. Moreover, we have shown that neither the c-fos promoter nor upstream sequences are absolute requirements for an efficient transcription arrest both in vivo and in vitro. Finally, we have characterized a 103-nucleotide-long intron 1 motif comprising the arrest site and sufficient for obtaining the block in a cell-free transcription assay.
Mechti
N
N
Laboratoire de Biochimie des Protéines, URA CNRS 1191, Génétique Moléculaire, Université des Sciences et Techniques du Languedoc, Montpellier, France.
Piechaczyk
M
M
Blanchard
J M
JM
Jeanteur
P
P
Lebleu
B
B
D000818:Animals; D001483:Base Sequence; D002151:Calorimetry; D002467:Cell Nucleus / Q000502:physiology; D002474:Cell-Free System; D015870:Gene Expression; D005786:Gene Expression Regulation; D007438:Introns; D007739:L Cells / Q000502:physiology; D007942:Leukemia, Experimental; D051379:Mice; D008961:Models, Structural; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes; D010957:Plasmids; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016760:Proto-Oncogene Proteins c-fos; D011519:Proto-Oncogenes; D015183:Restriction Mapping; D014158:Transcription, Genetic
D015345:Oligonucleotide Probes; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D011505:Protein-Tyrosine Kinases