The BRCA2 missense mutation K2497R suppressed self-degradation and increased ATP production and cell proliferation

Gerelmaa Enkhbat; Akira Nakanishi; Yoshio Miki

doi:10.1016/j.bbrc.2021.12.073

The BRCA2 missense mutation K2497R suppressed self-degradation and increased ATP production and cell proliferation

Biochem Biophys Res Commun. 2022 Jan 29:590:27-33. doi: 10.1016/j.bbrc.2021.12.073. Epub 2021 Dec 23.

Authors

Gerelmaa Enkhbat¹, Akira Nakanishi², Yoshio Miki³

Affiliations

¹ Department of Specialized Surgeries, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan; Department of Molecular Genetics, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.
² Department of Molecular Genetics, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan. Electronic address: nakanishi.mgen@mri.tmd.ac.jp.
³ Department of Molecular Genetics, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan; Department of Genetic Diagnosis, The Cancer Institute, Japanese Foundation for Cancer Research, 3-8-31 Ariake, Koto-ku, Tokyo, 135-8550, Japan. Electronic address: miki.mgen@mri.tmd.ac.jp.

PMID: 34968781
DOI: 10.1016/j.bbrc.2021.12.073

Abstract

Breast cancer susceptibility gene 2 (BRCA2) mediates genome maintenance during the S phase of the cell cycle, with important roles in replication stress, centrosome replication, and cytokinesis. In this study, we showed that a small heat shock protein, HSP27, interacted with and participated in the degradation of BRCA2 in estrogen-treated MCF-7 cells. BRCA2 degradation reportedly requires ubiquitination of the C-terminal region; thus, fragments of amino acid (aa) residues 2241-2940 were produced and assayed for their degradation following cycloheximide (CHX) treatment. The results showed that aa 2491-2580 affected the degradation of BRCA2, especially lysine (Lys) 2497. Furthermore, the K2497 A/R mutation increased ATP production and the proliferation of DLD-1 (BRCA2 knockout) cells compared to the cells expressing wild-type BRCA2-FLAG. Notably, a single residue, Lys2497, affected BRCA2 degradation, and K2497R is reportedly a missense mutation in hereditary breast cancer.

Keywords: BRCA2; Cell cycle; Cycloheximide; HSP27; K2497R; Ubiquitin.

Publication types

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

MeSH terms

Adenosine Triphosphate / biosynthesis*
Amino Acid Sequence
BRCA2 Protein / chemistry
BRCA2 Protein / genetics*
Breast Neoplasms / genetics
Breast Neoplasms / pathology
Cell Line, Tumor
Cell Proliferation / genetics
Female
HEK293 Cells
HSP27 Heat-Shock Proteins / metabolism
Humans
Lysine / genetics
Mutation, Missense / genetics*
Peptides / chemistry
Peptides / metabolism
Protein Binding
Proteolysis*
Ubiquitin / metabolism
Ubiquitination

Substances

BRCA2 Protein
BRCA2 protein, human
HSP27 Heat-Shock Proteins
Peptides
Ubiquitin
Adenosine Triphosphate
Lysine

Supplementary concepts

Breast Cancer, Familial