Cullin-3/KCTD10 complex is essential for K27-polyubiquitination of EIF3D in human hepatocellular carcinoma HepG2 cells

Masashi Maekawa; Hiromi Hiyoshi; Jun Nakayama; Kohki Kido; Tatsuya Sawasaki; Kentaro Semba; Eiji Kubota; Takashi Joh; Shigeki Higashiyama

doi:10.1016/j.bbrc.2019.07.010

Cullin-3/KCTD10 complex is essential for K27-polyubiquitination of EIF3D in human hepatocellular carcinoma HepG2 cells

Biochem Biophys Res Commun. 2019 Sep 3;516(4):1116-1122. doi: 10.1016/j.bbrc.2019.07.010. Epub 2019 Jul 5.

Authors

Masashi Maekawa¹, Hiromi Hiyoshi², Jun Nakayama³, Kohki Kido⁴, Tatsuya Sawasaki⁴, Kentaro Semba³, Eiji Kubota², Takashi Joh⁵, Shigeki Higashiyama⁶

Affiliations

¹ Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Shitsukawa, Toon, Ehime, 791-0295, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Shitsukawa, Toon, Ehime, 791-0295, Japan. Electronic address: masashim@m.ehime-u.ac.jp.
² Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, Aichi, 467-8601, Japan.
³ Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, 2-2 Wakamatsucho, Shinjuku-ku, Tokyo, 162-8480, Japan.
⁴ Division of Cell-Free Science, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
⁵ Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, Aichi, 467-8601, Japan; Gamagori City Hospital, 1-1 Mukaida, Hirata-Cho, Gamagori, Aichi, 443-8501, Japan.
⁶ Division of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime University, Shitsukawa, Toon, Ehime, 791-0295, Japan; Department of Biochemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Shitsukawa, Toon, Ehime, 791-0295, Japan. Electronic address: shigeki@m.ehime-u.ac.jp.

PMID: 31280863
DOI: 10.1016/j.bbrc.2019.07.010

Abstract

Eukaryotic translation initiation factor 3 subunit D (EIF3D) binds to the 5'-cap of specific mRNAs, initiating their translation into polypeptides. From a pathological standpoint, EIF3D has been observed to be essential for cell growth in various cancer types, and cancer patients with high EIF3D mRNA levels exhibit poor prognosis, indicating involvement of EIF3D in oncogenesis. In this study, we found, by mass spectrometry, that Cullin-3 (CUL3)/KCTD10 ubiquitin (Ub) ligase forms a complex with EIF3D. We also demonstrated that EIF3D is K27-polyubiquitinated at the lysine 153 and 275 residues in a KCTD10-dependent manner in human hepatocellular carcinoma HepG2 cells. Similar to other cancers, high expression of EIF3D significantly correlated with poor prognosis in hepatocellular carcinoma patients, and depletion of EIF3D drastically suppressed HepG2 cell proliferation. These results indicate that EIF3D is a novel substrate of CUL3/KCTD10 Ub ligase and suggest involvement of K27-polyubiquitinated EIF3D in the development of hepatocellular carcinoma.

Keywords: Cullin-3 (CUL3); EIF3D; Hepatocellular carcinoma; KCTD10.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carcinoma, Hepatocellular / metabolism*
Cullin Proteins / metabolism*
Eukaryotic Initiation Factor-3 / metabolism*
Hep G2 Cells
Humans
Liver Neoplasms / metabolism*
Potassium Channels, Voltage-Gated / metabolism*
Protein Interaction Maps
Ubiquitination

Substances

CUL3 protein, human
Cullin Proteins
EIF3D protein, human
Eukaryotic Initiation Factor-3
KCTD10 protein, human
Potassium Channels, Voltage-Gated