Identification of 3',5'-cyclic adenosine monophosphate-inducible nuclear factors binding to the human urokinase promoter in mouse Sertoli cells.
Transcription factors which recognize both the SV40 promoter and the proximal promoter region of the human urokinase-type plasminogen activator (h-uPA) gene are present in nuclear extracts from primary cultures of mouse Sertoli cells; prolonged (more than 12 h) (Bu)2cAMP stimulation of Sertoli cells induces the formation of different specific DNA-protein complexes. A discrete region in the h-uPA promoter, between -54 and -42, is essential for the formation of the cAMP-induced DNA-protein complexes. Mutation of the sequence between -54 and -42 abolishes the response to cAMP of the proximal h-uPA promoter in Sertoli cells. A protein, recognizing a sequence centered around the GC-box present between -48 and -43, is detected by Southwestern analysis, and it is clearly induced by (Bu)2cAMP stimulation. Interaction between this protein and a second factor, recognizing a purine-rich sequence between -53 and -46, partially overlapping the GC-box, is needed for the formation of the cAMP-induced DNA-protein complexes. A preformed complex between the cAMP-induced GC-box-binding factor and the second factor can be detected using nondenaturing conditions during Southwestern analysis.
Grimaldi
P
P
Department of Public Health and Cell Biology, University of Rome Tor Vergata, Italy.
Piscitelli
D
D
Albanesi
C
C
Blasi
F
F
Geremia
R
R
Rossi
P
P
D000818:Animals; D001483:Base Sequence; D003994:Bucladesine / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D015183:Restriction Mapping; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism*; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics*
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D003994:Bucladesine; D000242:Cyclic AMP; D015500:Chloramphenicol O-Acetyltransferase; D014568:Urokinase-Type Plasminogen Activator