Novel upstream elements and the TATA-box region mediate preferential transcription from the uteroglobin promoter in endometrial cells.
To understand the mechanisms responsible for endometrium-specific expression of the uteroglobin gene, we have compared transcription from the uteroglobin promoter in a human endometrial cell line (Ishikawa) and in HeLa cells. In transient transfection experiments and in nuclear extracts, sequences from -395 to +14 of the uteroglobin gene are able to promote transcription of a reporter gene more efficiently in Ishikawa cells than in HeLa cells relative to the RSV or the SV40 early promoter. Analysis of progressive 5'-deletion mutants identifies three promoter regions, -258/-220, -205/-177, and -96/-35, that are important for preferential transcription in endometrial cells. DNase I footprinting experiments with nuclear extracts from Ishikawa and HeLa cells reveal a series of defined protections overlapping these regions. The relative intensity of individual protections differs between the two cell lines. Oligonucleotide competition experiments suggest that similar factor(s) bind(s) to the two relevant upstream regions of the promoter that share no homology to known regulatory elements. A protection over the TATA-box is detected only with extracts from Ishikawa cells. Band shift experiments show that an Ishikawa-specific factor binds to sequences overlapping the TATA-box region that are partially conserved in other endometrium-expressed genes. We propose that novel transcription factors mediate endometrium-specific expression of the uteroglobin gene in conjunction with a tissue-specific factor that binds to the TATA-box region.
Misseyanni
A
A
Institut für Molekularbiologie und Tumorforschung (IMT), Marburg, FRG.
Klug
J
J
Suske
G
G
Beato
M
M
D001483:Base Sequence; D001667:Binding, Competitive; D002474:Cell-Free System; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis; D016172:DNA Fingerprinting; D004717:Endometrium / Q000166:cytology / Q000378:metabolism*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012313:RNA / Q000032:analysis; D016385:TATA Box; D014158:Transcription, Genetic; D014162:Transfection; D014598:Uteroglobin / Q000235:genetics*
D009841:Oligonucleotides; D012313:RNA; D014598:Uteroglobin; D015500:Chloramphenicol O-Acetyltransferase