Protein phosphatase 2A regulated by USP7 is polyubiquitinated and polyneddylated

Do,Hyeon-Ah; Baek,Kwang-Hyun

doi:10.3892/or.2022.8335

Protein phosphatase 2A regulated by USP7 is polyubiquitinated and polyneddylated

Authors:
- Hyeon-Ah Do
- Kwang-Hyun Baek
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Affiliations: Department of Biomedical Science, CHA University, Seongnam‑si, Gyeonggi‑do 13488, Republic of Korea
Published online on: May 19, 2022 https://doi.org/10.3892/or.2022.8335
Article Number: 124

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Abstract

Ubiquitin‑specific protease 7 (USP7) participates in the ubiquitin‑proteasome system (UPS), and is considered an essential regulator of substrate stability in cancers. In a previous study, the substrates that bind to USP7 were separated through two‑dimensional electrophoresis (2‑DE), which resulted in the identification of protein phosphatase 2A (PP2A) through matrix‑assisted laser desorption‑ionization time‑of‑flight mass spectrometry (MALDI‑TOF/MS) analysis. In the present study, GST pull‑down assay was performed to determine whether USP7 and PP2A directly bind to each other. Immunocytochemistry assay confirmed that USP7 co‑localizes with PP2A in the cytoplasm and nucleus of HeLa cells. Moreover, western blotting and immunoprecipitation were performed to determine whether polyubiquitination and polyneddylation of PP2A were formed. The results of the present study demonstrated that USP7 was a deubiquitinating enzyme of PP2A, and regulated the ubiquitination and stability of PP2A through the K48‑linked polyubiquitin chains. Consequently, the knockdown of USP7 reduced the expression of PP2A. The data of the present study revealed the cellular association between USP7 and PP2A, a new substrate of USP7.

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