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Updated: Jun 25, 2026

In Vitro Ubiquitination and Deubiquitination Assays of Nucleosomal Histones
Published on: July 25, 2019
Nucleolar structure and function are regulated by the deubiquitylating enzyme USP36.
Akinori Endo1, Masaki Matsumoto, Toshifumi Inada
1Department of Biological Sciences, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
USP36, a deubiquitylating enzyme, regulates nucleolar activity by stabilizing key proteins like nucleophosmin/B23. Its depletion impairs ribosome biogenesis and cell proliferation.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- The nucleolus is essential for ribosome biogenesis.
- Protein ubiquitylation is implicated in nucleolar functions.
- The deubiquitylating enzyme USP36's role in the nucleolus was unknown.
Purpose of the Study:
- To investigate the function of USP36 in mammalian cells.
- To determine USP36's role in nucleolar activity and ribosome biogenesis.
Main Methods:
- Localization studies of USP36 in nucleoli.
- Dominant-negative inhibition and RNA interference (RNAi) to assess USP36 function.
- Analysis of nucleolar protein ubiquitylation and stability.
- Assessment of rRNA transcription, processing, and cell proliferation.
Main Results:
- USP36 localizes to the nucleolus and deubiquitylates nucleolar proteins nucleophosmin/B23 and fibrillarin.
- USP36 stabilizes these proteins by preventing ubiquitylation-mediated degradation.
- USP36 depletion reduces rRNA synthesis, impairs nucleolar morphology, and decreases cell proliferation.
Conclusions:
- USP36 is a crucial regulator of nucleolar structure and function.
- USP36 maintains nucleolar integrity and ribosome biogenesis through deubiquitylation of key substrates.
- USP36 activity is vital for cellular growth and proliferation.
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