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The Ubiquitin-specific Protease USP36 Associates with the Microprocessor Complex and Regulates miRNA Biogenesis by

Yanping Li1, Timothy S Carey1, Catherine H Feng1

  • 1Department of Molecular and Medical Genetics, School of Medicine, Oregon Health & Science University, Portland, Oregon.

Cancer Research Communications
|March 23, 2023
PubMed

Insights

USP36 regulates miRNA biogenesis by SUMOylating DGCR8, a key protein in the process. This finding is significant as USP36 overexpression is linked to cancer development.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • MicroRNA (miRNA) biogenesis is crucial for gene regulation and is tightly controlled by protein modifications.
  • Aberrant miRNA biogenesis is implicated in human diseases, including cancer.
  • DGCR8, a component of the microprocessor complex, is known to undergo SUMOylation, but the responsible enzyme was unidentified.

Purpose of the Study:

  • To identify the SUMO ligase responsible for DGCR8 SUMOylation.
  • To elucidate the role of USP36 in miRNA biogenesis.
  • To investigate the functional consequences of DGCR8 SUMOylation and USP36 activity in cellular processes like proliferation.

Main Methods:

  • Co-immunoprecipitation assays to detect protein interactions.
  • Western blotting to assess protein modification (SUMOylation) and levels.
  • RNA immunoprecipitation (RIP) to study pri-miRNA binding.
  • Cell proliferation assays.
  • Knockdown and overexpression studies of USP36 and DGCR8.

Main Results:

  • USP36 was identified as a novel regulator of DGCR8, interacting with the microprocessor complex.
  • USP36 promotes DGCR8 SUMOylation, specifically by SUMO2, enhancing DGCR8 binding to pri-miRNAs.
  • Knockdown of USP36 impaired pri-miRNA processing and reduced mature miRNA levels.
  • A SUMOylation-defective DGCR8 mutant inhibited cell proliferation.

Conclusions:

  • USP36 is a critical enzyme for miRNA biogenesis through its regulation of DGCR8 SUMOylation.
  • The USP36-DGCR8 pathway represents a potential target in cancer therapy due to USP36's frequent overexpression in tumors.

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