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Purification of Ubiquitinated p53 Proteins from Mammalian Cells
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Ovarian tumor domain-containing protein 1 deubiquitinates and stabilizes p53

Shudong Piao1, Han Zhong Pei1, Bin Huang1

  • 1Department of Biochemistry and Molecular Biology, College of Medicine, Yeungnam University, South Korea.

Cellular Signalling
|February 21, 2017
PubMed

Insights

OTU domain-containing protein 1 (OTUD1) deubiquitinates and stabilizes p53, a key tumor suppressor. This novel regulator enhances p53 activity, suppresses tumor growth, and promotes apoptosis, offering new therapeutic avenues.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Protein Biochemistry

Background:

  • Ubiquitination and deubiquitination are critical for regulating p53 protein stability and activity.
  • Deubiquitinases (DUBs) stabilize p53, while E3 ubiquitin ligases destabilize it.
  • The role of OTU domain-containing protein 1 (OTUD1) in p53 regulation was previously unknown.

Purpose of the Study:

  • To identify novel deubiquitinases (DUBs) that stabilize p53.
  • To investigate the role of OTU domain-containing protein 1 (OTUD1) in p53 stability and activity.

Main Methods:

  • Screening of ovarian tumor (OTU) subfamily proteins.
  • Overexpression and knockdown systems in cells.
  • Co-immunoprecipitation to assess protein interactions.
  • Analysis of p53 target gene expression (p21, Mdm2).
  • Colony formation assays.
  • Apoptosis assays (caspase-3 and PARP cleavage).

Main Results:

  • OTUD1 was identified as a novel DUB that directly interacts with and deubiquitinates p53.
  • OTUD1 overexpression increased p53 stability and activity, while OTUD1 knockdown decreased them.
  • Functional OTUD1, but not an inactive mutant, stabilized p53, upregulated p21 and Mdm2, suppressed colony formation, and induced apoptosis.
  • OTUD1's deubiquitinating activity is essential for p53 stabilization.

Conclusions:

  • OTUD1 is a novel regulator of p53 stability and activity.
  • OTUD1 functions as a DUB to stabilize p53, thereby suppressing tumor cell proliferation and promoting apoptosis.
  • OTUD1 represents a potential therapeutic target in cancer treatment.
Keywords:
Cell growthDeubiquitinaseOvarian tumor domain-containing protein 1Ubiquitinp53

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