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PACT/PRKRA and p53 regulate transcriptional activity of DMRT1

Kazuko Fujitani1, Asako Otomo2, Yuto Nagayama3

  • 1Kitasato University, Gene Analysis Center, School of Medicine, Sagamihara, Japan.

Genetics and Molecular Biology
|April 7, 2020
PubMed

Insights

PACT/PRKRA protein enhances doublesex and mab-3 related transcription factor 1 (DMRT1) activity, while p53 represses it. This interaction is crucial for germline stem cell fate in Xenopus laevis.

Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • The transcription factor DMRT1 plays key roles in somatic-cell masculinization and germ-cell development in vertebrates.
  • The precise transcriptional regulation of DMRT1, particularly in germline development, remains largely uncharacterized.

Purpose of the Study:

  • To identify DMRT1-interacting proteins in Xenopus laevis testes.
  • To investigate the regulatory effects of identified interacting proteins, specifically PACT/PRKRA and p53, on DMRT1 transcriptional activity.

Main Methods:

  • Immunoprecipitation using an anti-DMRT1 antibody followed by MS/MS analysis to identify interacting proteins.
  • Luciferase reporter assays in transfected 293T cells to assess DMRT1 transcriptional activity.
  • In situ hybridization to analyze Pact/Prkra mRNA expression patterns in tadpole gonads.

Main Results:

  • PACT/PRKRA was identified as a DMRT1-interacting protein in Xenopus laevis testes.
  • PACT/PRKRA significantly enhanced DMRT1-driven luciferase activity, whereas p53 significantly repressed it.
  • p53 attenuated the PACT/PRKRA-mediated enhancement of DMRT1 activity.
  • Pact/Prkra mRNA showed high expression in both male and female germline stem cells of tadpoles.

Conclusions:

  • PACT/PRKRA may act as a positive regulator, and p53 as a negative regulator, of DMRT1 activity.
  • These findings suggest a novel regulatory mechanism involving PACT/PRKRA and p53 in controlling DMRT1 function for germline stem cell fate determination.

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