β-Catenin (CTNNB1) in the Mouse Uterus During Decidualization and the Potential Role of Two Pathways in Regulating Its Degradation

β-catenin plays a role in cell adhesion and as a transcriptional coactivator. Its levels are regulated in cells by controlling its degradation through ubiquitination by two different E3 ligase complexes. One complex contains β-transducing repeat containing (BTRC) protein, which binds to β-catenin when phosphorylated on specific (S33 and S37) residues, whereas the other involves calcyclin-binding protein (CACYBP). The aim of this study was to determine the localization and levels of total and active (S33/S37-dephosphorylated) β-catenin in the pregnant mouse uteri and those undergoing artificially stimulated decidualization. These two forms of β-catenin were localized almost exclusively to the endometrial epithelia just prior to the onset of implantation. Although this localization continued after the onset of implantation, there were less epithelial cells present in areas of the uterus undergoing decidualization. Rather, there was a progressive increase in β-catenin localization in endometrial stromal cells undergoing decidualization in the anti-mesometrial and, to a lesser extent, in the mesometrial regions. The presence of a conceptus was not required for the changes in localization seen in the pregnant uterus because similar findings were also seen in uteri undergoing artificially stimulated decidualization. Finally, overall levels of total, active (S33 and S37 dephosphorylated), and phosphorylated (S33/S37/T42) β-catenin protein and the steady-state levels of calcyclin-binding protein mRNA changed in the uterus during decidualization. The result of this study shows the changing localization and levels of β-catenin in the mouse uterus during decidualization. Further, the results suggest potential roles for both the BTRC and CACYBP E3 ligase mechanisms of β-catenin ubiquitination in the uterus during decidualization.

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COUP-TFII Regulates Human Endometrial Stromal Genes Involved in Inflammation

Molecular Endocrinology ◽

10.1210/me.2013-1191 ◽

2013 ◽

Vol 27 (12) ◽

pp. 2041-2054 ◽

Cited By ~ 32

Author(s):

Xilong Li ◽

Michael J. Large ◽

Chad J. Creighton ◽

Rainer B. Lanz ◽

Jae-Wook Jeong ◽

...

Keyword(s):

Cell Fate ◽

Cell Biology ◽

Embryo Implantation ◽

Orphan Nuclear Receptor ◽

Loss Of Function ◽

Mouse Uterus ◽

Endometrial Stromal Cells ◽

Endometrial Stroma ◽

Stroma Cell

Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII; NR2F2) is an orphan nuclear receptor involved in cell-fate specification, organogenesis, angiogenesis, and metabolism. Ablation of COUP-TFII in the mouse uterus causes infertility due to defects in embryo attachment and impaired uterine stromal cell decidualization. Although the function of COUP-TFII in uterine decidualization has been described in mice, its role in the human uterus remains unknown. We observed that, as in mice, COUP-TFII is robustly expressed in the endometrial stroma of healthy women, and its expression is reduced in the ectopic lesions of women with endometriosis. To interrogate the role of COUP-TFII in human endometrial function, we used a small interfering RNA-mediated loss of function approach in primary human endometrial stromal cells. Attenuation of COUP-TFII expression did not completely block decidualization; rather it had a selective effect on gene expression. To better elucidate the role of COUP-TFII in endometrial stroma cell biology, the COUP-TFII transcriptome was defined by pairing microarray comparison with chromatin immunoprecipitation followed by deep sequencing. Gene ontology analysis demonstrates that COUP-TFII regulates a subset of genes in endometrial stroma cell decidualization such as those involved in cell adhesion, angiogenesis, and inflammation. Importantly this analysis shows that COUP-TFII plays a role in controlling the expression of inflammatory cytokines. The determination that COUP-TFII plays a role in inflammation may add insight into the role of COUP-TFII in embryo implantation and in endometrial diseases such as endometriosis.

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A Genomic Approach to Identify Novel Progesterone Receptor Regulated Pathways in the Uterus during Implantation

Molecular Endocrinology ◽

10.1210/me.2002-0270 ◽

2002 ◽

Vol 16 (12) ◽

pp. 2853-2871 ◽

Cited By ~ 87

Author(s):

Yong-Pil Cheon ◽

Quanxi Li ◽

Xueping Xu ◽

Francesco J. DeMayo ◽

Indrani C. Bagchi ◽

...

Keyword(s):

Gene Networks ◽

Time Course ◽

Target Genes ◽

Reproductive Tract ◽

Cell Types ◽

Temporal Analysis ◽

Pregnant Mouse ◽

Initial Examination ◽

Mouse Uterus ◽

Pregnant Uterus

Abstract The cellular actions of steroid hormone progesterone (P) are mediated via its nuclear receptors, which regulate the expression of specific target genes. The identity of gene networks that are regulated by the P receptors (PRs) in the uterus at various stages of the reproductive cycle and pregnancy, however, remain largely unknown. In this study, we have used oligonucleotide microarrays to identify mRNAs whose expression in the pregnant mouse uterus is modulated by RU486, a well-characterized PR antagonist, which is also an effective inhibitor of implantation. We found that, in response to RU486, expression of mRNAs corresponding to 78 known genes was down-regulated at least 2-fold in the preimplantation mouse uterus. The PR regulation of several of these genes was ascertained by administering P to ovariectomized wild-type and PR knockout (PRKO) mice. Detailed spatio-temporal analysis of these genes in the pregnant uterus indicated that their expression in the epithelium and stroma could be correlated with the expression of PR in those cell types. Furthermore, time-course studies suggested that many of these genes are likely primary targets of PR regulation. We also identified 70 known genes that were up-regulated at least 2-fold in the pregnant uterus in response to RU486. Interestingly, initial examination of a number of RU486-inducible genes reveals that their uterine expression is also regulated by estrogen. The identification of several novel PR-regulated gene pathways in the reproductive tract is an important step toward understanding how P regulates the physiological events leading to implantation.

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Expression of corticotrophin-releasing hormone in the mouse uterus: participation in embryo implantation

Journal of Endocrinology ◽

10.1677/joe.0.1630221 ◽

1999 ◽

Vol 163 (2) ◽

pp. 221-227 ◽

Cited By ~ 18

Author(s):

I Athanassakis ◽

V Farmakiotis ◽

I Aifantis ◽

A Gravanis ◽

S Vassiliadis

Keyword(s):

Cell Culture ◽

Embryo Implantation ◽

Mouse Uterus ◽

Pregnant Uterus ◽

Cell Culture Techniques ◽

Releasing Hormone ◽

Culture Techniques ◽

Blastocyst Implantation ◽

Implantation Sites

The detection of corticotropin-releasing hormone (CRH) in the pregnant and non-pregnant uterus has driven research to determine the role of this 41 amino acid neuropeptide in the female reproductive system. As concentrations of CRH mRNA and its peptide product are greater in the implantation sites of the early pregnant uterus compared with the regions between implantation sites, CRH has been hypothesised to participate in blastocyst implantation. Using the mouse system as an experimental model, we studied the distribution of CRH in the uterus during the oestrus cycle and early gestational period, and now provide evidence for its involvement in embryo implantation using cell culture techniques. The percentage of CRH-positive uterine cells and the amount of CRH released during anoestrus, pro-oestrus and oestrus were determined by immunofluorescence and ELISA experiments respectively. The highest number of intracellularly CRH-positive cells was obtained during pro-oestrus, whereas the highest CRH concentration in uterine cell culture supernatants was detected during anoestrus. At early stages of gestation, CRH was detected in the endometrium on days 2, 3 and 4 of pregnancy and in the myometrium on days 3 and 4, whereas it was undetectable on day 5. The functional role of CRH during early gestation was evaluated by administering anti-CRH antibody to mice from day 3 to day 8 of pregnancy. This treatment resulted in implantation failure in 60% of the cases, in which implantation sites, although clearly present in the uterus, had failed to host an embryo. These results provide direct evidence about the involvement of CRH in murine embryo implantation and are in agreement with hypotheses postulated in humans.

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Tethering of an E3 ligase by PCM1 regulates the abundance of centrosomal KIAA0586/Talpid3 and promotes ciliogenesis

eLife ◽

10.7554/elife.12950 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 55

Author(s):

Lei Wang ◽

Kwanwoo Lee ◽

Ryan Malonis ◽

Irma Sanchez ◽

Brian D Dynlacht

Keyword(s):

Binding Protein ◽

E3 Ligase ◽

Human Cells ◽

Gene Encoding ◽

Amino Terminal ◽

Terminal Domain ◽

Integral Component ◽

Specific Proteins ◽

Amino Terminal Domain

To elucidate the role of centriolar satellites in ciliogenesis, we deleted the gene encoding the PCM1 protein, an integral component of satellites. PCM1 null human cells show marked defects in ciliogenesis, precipitated by the loss of specific proteins from satellites and their relocation to centrioles. We find that an amino-terminal domain of PCM1 can restore ciliogenesis and satellite localization of certain proteins, but not others, pinpointing unique roles for PCM1 and a group of satellite proteins in cilium assembly. Remarkably, we find that PCM1 is essential for tethering the E3 ligase, Mindbomb1 (Mib1), to satellites. In the absence of PCM1, Mib1 destabilizes Talpid3 through poly-ubiquitylation and suppresses cilium assembly. Loss of PCM1 blocks ciliogenesis by abrogating recruitment of ciliary vesicles associated with the Talpid3-binding protein, Rab8, which can be reversed by inactivating Mib1. Thus, PCM1 promotes ciliogenesis by tethering a key E3 ligase to satellites and restricting it from centrioles.

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A cDNA cloned from pregnant mouse uterus exhibits temporo-spatial expression and predicts a novel protein

Biochemical Journal ◽

10.1042/bj3300947 ◽

1998 ◽

Vol 330 (2) ◽

pp. 947-950 ◽

Cited By ~ 16

Author(s):

W. John KASIK

Keyword(s):

Late Pregnancy ◽

Pregnant Mouse ◽

Open Reading Frame ◽

Mouse Uterus ◽

Pregnant Uterus ◽

Reading Frame ◽

Spatial Expression ◽

Conceptual Translation ◽

Novel Protein ◽

Specific Mrna

A cDNA was cloned from a pregnant mouse uterus cDNA library. On conceptual translation, the cDNA has one long open reading frame that predicts a novel protein of 606 amino acids. This protein is principally composed of two CUB domains and a ZP domain; motifs found in proteins implicated in egg-sperm recognition. Probes derived from the cDNA were used to conduct Northern hybridizations. The expression of this mRNA is temporal; message first appears in the uterus 6 days prior to birth, it increases each subsequent day to attain maximal levels at 3 days prior to birth and then abruptly decreases during the last 3 days of pregnancy. The expression of this mRNA is restricted; message is abundant in the uterus during late pregnancy, but it is not found in non-pregnant uterus or in a variety of adult or fetal tissues. The temporo-spatial expression of this pregnant uterus specific mRNA and the consolidation in the predicted protein of two motifs implicated in early pregnancy events suggests that the product of the gene represented by this mRNA may play an important role in events that transpire during late pregnancy.

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Protein Inhibitor of Activated STAT 1 (PIAS1) Is Identified as the SUMO E3 Ligase of CCAAT/Enhancer-Binding Protein β (C/EBPβ) during Adipogenesis

Molecular and Cellular Biology ◽

10.1128/mcb.00723-13 ◽

2013 ◽

Vol 33 (22) ◽

pp. 4606-4617 ◽

Cited By ~ 31

Author(s):

Yang Liu ◽

Ya-Dong Zhang ◽

Liang Guo ◽

Hai-Yan Huang ◽

Hao Zhu ◽

...

Keyword(s):

Adipocyte Differentiation ◽

Binding Protein ◽

E3 Ligase ◽

Inhibitory Effect ◽

Cellular Processes ◽

Enhancer Binding Protein ◽

Ccaat Enhancer Binding Protein ◽

Sumo E3 Ligase ◽

Attachment Factor

It is well recognized that PIAS1, a SUMO (small ubiquitin-like modifier) E3 ligase, modulates such cellular processes as cell proliferation, DNA damage responses, and inflammation responses. Recent studies have shown that PIAS1 also plays a part in cell differentiation. However, the role of PIAS1 in adipocyte differentiation remains unknown. CCAAT/enhancer-binding protein β (C/EBPβ), a major regulator of adipogenesis, is a target of SUMOylation, but the E3 ligase responsible for the SUMOylation of C/EBPβ has not been identified. The present study showed that PIAS1 functions as a SUMO E3 ligase of C/EBPβ to regulate adipogenesis. PIAS1 expression was significantly and transiently induced on day 4 of 3T3-L1 adipocyte differentiation, when C/EBPβ began to decline. PIAS1 was found to interact with C/EBPβ through the SAP (scaffold attachment factor A/B/acinus/PIAS) domain and SUMOylate it, leading to increased ubiquitination and degradation of C/EBPβ. C/EBPβ became more stable when PIAS1 was silenced by RNA interference (RNAi). Moreover, adipogenesis was inhibited by overexpression of wild-type PIAS1 and promoted by knockdown of PIAS1. The mutational study indicated that the catalytic activity of SUMO E3 ligase was required for PIAS1 to restrain adipogenesis. Importantly, the inhibitory effect of PIAS1 overexpression on adipogenesis was rescued by overexpressed C/EBPβ. Thus, PIAS1 could play a dynamic role in adipogenesis by promoting the SUMOylation of C/EBPβ.

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Regulation and Function of Laminin A5 during Mouse and Human Decidualization

International Journal of Molecular Sciences ◽

10.3390/ijms23010199 ◽

2021 ◽

Vol 23 (1) ◽

pp. 199

Author(s):

Zhen-Shan Yang ◽

Hai-Yang Pan ◽

Wen-Wen Shi ◽

Si-Ting Chen ◽

Ying Wang ◽

...

Keyword(s):

Early Pregnancy ◽

Stromal Cells ◽

Basement Membranes ◽

Pivotal Role ◽

Cellular Phenotype ◽

Mouse Uterus ◽

Endometrial Stromal Cells ◽

And Function

Decidualization is essential to the establishment of pregnancy in rodents and primates. Laminin A5 (encoding by Laminin α5) is a member of the laminin family, which is mainly expressed in the basement membranes. Although laminins regulate cellular phenotype maintenance, adhesion, migration, growth, and differentiation, the expression, function, and regulation of laminin A5 during early pregnancy are still unknown. Therefore, we investigated the expression and role of laminin A5 during mouse and human decidualization. Laminin A5 is highly expressed in mouse decidua and artificially induced deciduoma. Laminin A5 is significantly increased under in vitro decidualization. Laminin A5 knockdown significantly inhibits the expression of Prl8a2, a marker for mouse decidualization. Progesterone stimulates the expression of laminin A5 in ovariectomized mouse uterus and cultured mouse stromal cells. We also show that progesterone regulates laminin A5 through the PKA-CREB-C/EBPβ pathway. Laminin A5 is also highly expressed in human pregnant decidua and cultured human endometrial stromal cells during in vitro decidualization. Laminin A5 knockdown by siRNA inhibits human in vitro decidualization. Collectively, our study reveals that laminin A5 may play a pivotal role during mouse and human decidualization via the PKA-CREB-C/EBPβ pathway.

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