Functional withdrawal of progesterone and the initiation of labour.

摘要

The onset of labour interplays factors that originate from both mother and fetus. It involves the transition of myometrial cell phenotype from uterine quiescence to uterine activation and stimulation of uterine contractions. It is characterized by a change in expression of a set of myometrium contraction associated proteins (CAP), which activate the myometrium in preparation for stimulation by uterotonins to initiate labour. Progesterone plays an essential role in signalling transduction to modulate the function of these CAP. Understanding the mechanisms that control CAP gene expression would benefit the development of effective means of preventing preterm delivery and the consequent neonatal mortality and morbidity. The objective of this thesis is to explore the switch of progesterone signalling from an active state through most of the pregnant stages to a functional withdrawal at term. My studies confirmed that the progesterone receptor-B (PRB) is a strong transcriptional activator, while PRA can antagonize PRB in the context of the myometrial cell. The molecular mechanism underlying the function of PRB involves two LXXLL motifs that are lacking in PRA. These motifs mediate intramolecular protein interactions between the AF3 (Activation Function 3) domain and the C-terminus of PR in a ligand dependent manner. The functional consequence of this interaction is further enhanced by the presence of coactivators such as GRIP-1. I have also isolated a novel PR interacting protein, PSF, previously identified as a pre-mRNA splicing factor. The interaction between PR and PSF is confirmed by both in vivo and in vitro protein assays and the interacting sites are located in the AF3 and the DBD (DNA Binding Domain) of PR and in the RRM II (RNA Recognition Motif II) of PSF. PSF is shown to inhibit transactivation of PR in several cellular promoter contexts. I provide evidence that the corepression of PR by PSF involves multiple mechanisms including the enhancement of PR protein degradation and interference of PR binding to PRE (progesterone response element). I have also located two regions within PSF that possess inhibitory functions. Most importantly, I have demonstrated that upregulation of PSF expression in rat myometrium at term is temporally correlated with the deregulation of PR protein. These data collectively support a role for PSF as a critical corepressor that contributes to the functional withdrawal of progesterone at term labour.