Definition of a negative modulation domain in the human progesterone receptor.
The progesterone receptor (PR) occurs in two major forms, the full-length PRB and the amino-truncated PRA, which lacks 164 amino-terminal residues. PRB functions as a strong transcriptional activator of progesterone-responsive genes, whereas PRA is inactive in several cell types where it may even act as a trans-dominant repressor of PRB and other steroid receptors, like the glucocorticoid receptor or, reportedly, the estrogen receptor. We initially observed that a PR deleted of its entire amino domain (PR538-C) is incapable of trans-repressing PRB or glucocorticoid receptor, suggesting that a negative modulation domain must be contained in the region between position 165 and 538. After testing progressive deletion mutants and chimeras, we demonstrate that this negative modulating domain is confined within 120 residues in the amino-terminal region and that it contains a subdomain of 40 residues that is crucial for intermolecular transrepression. Duplication, deletion, and transplantation of the negative modulation domain show that the negative modulation domain has only a limited functional autonomy. In our hands, transrepression of estrogen receptor could not be substantiated, and, under our conditions, at least an equimolar concentration of PRA expression plasmid is required for transrepression. Our deletion studies reveal domains that correlate with strong homology patches between the amino-terminal domains of mammalian and avian PR.
Huse
B
B
Biochemistry Institute, Université de Fribourg, Pérolles, Switzerland.
Verca
S B
SB
Matthey
P
P
Rusconi
S
S
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002645:Chickens; D004247:DNA / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D011374:Progesterone / Q000502:physiology; D011980:Receptors, Progesterone / Q000502:physiology*; D016415:Sequence Alignment; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation; D014162:Transfection
D011980:Receptors, Progesterone; D011374:Progesterone; D004247:DNA