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Nucleic Acids Research, 1988, Vol. 16, No. 12 5459-5472
© 1988

Articles

Organisation of the entire rabbit progesterone receptor mRNA and of the promoter and 5' flanking region of the gene

Micheline Misrahi, Hugues Loosfelt, Michel Atger, Cécile Mériel, Véronique Zerah, Philippe Dessen1 and Edwin Milgrom*

INSERM U.135, Faculté de Médicine Paris-Sud 94275 Le Kremlin-Bicêtre Cedex, France 1Laboratoire de Biochimie (UA 240 CNRS), Ecole Polytechnique 91128 Palaiseau, France

*To whom correspondence should be addressed

Received February 19, 1988. Revised May 18, 1988. Accepted May 18, 1988.

cDNA clones corresponding to the 3' and 5' non coding regions of the rabbit progesterone receptor (rPR) mRNA and genomic clones corresponding to the promoter and 5' flanking region of this gene were isolated and sequenced up to nucleotide -2761. The 3' non coding region is very long (3058–3553 nucleotides) and contains three different polyadenylation sites. Primer extension experiments and S1 mapping showed the existence of 2 transcription initiation sites 699 and 712 bp upstream from the Initiator ATG. The promoter region contains two modified TATA boxes: TAGAAA at -17 and TAGA at -37bp. A CAACT sequence is present at position -100 and one consensus binding site for the transcription factor Sp1 is found at position -51. A 317 bp sequence was observed (positions -2590 to -2273) which belongs to the C family of the short interspersed repeats of the rabbit. Sequences resembling the consensus for estrogen and progesterone responsive elements are observed at several locations in the 5' flanking region. The progesterone receptor is present in tissue extracts mainly as a mixture of two molecular species (110 and 79 kDa) whose origin remains currently debated. By Northern blot analysis we have shown, using rabbit and human mRNAs, that these receptor species are not derived from separate mRNAs. Transcriptlon-translatlon experiments also showed that, at least in vitro, they are not derived by use of different translation initiation sites on the same messenger RNA.


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