Clathrin heavy chain phosphorylated at T606 plays a role in proper cell division

Yusuke Yabuno; Toshihiro Uchihashi; Towa Sasakura; Hiroyuki Shimizu; Yoko Naito; Kohshiro Fukushima; Kaori Ota; Mikihiko Kogo; Hiroshi Nojima; Norikazu Yabuta

doi:10.1080/15384101.2019.1637201

Clathrin heavy chain phosphorylated at T606 plays a role in proper cell division

Cell Cycle. 2019 Aug;18(16):1976-1994. doi: 10.1080/15384101.2019.1637201. Epub 2019 Jul 4.

Authors

Affiliations

¹ a First Department of Oral and Maxillofacial Surgery, Graduate School of Dentistry, Osaka University , Osaka , Japan.
² b Department of Molecular Genetics, Research Institute for Microbial Diseases, Osaka University , Osaka , Japan.
³ c Division of Cancer Cell Regulation, Aichi Cancer Center Research Institute , Aichi , Japan.
⁴ d Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University , Suita , Osaka , Japan.

Abstract

Clathrin regulates mitotic progression, in addition to membrane trafficking. However, the detailed regulatory mechanisms of clathrin during mitosis remain elusive. Here, we demonstrate novel regulation of clathrin during mitotic phase of the cell cycle. Clathrin heavy chain (CHC) was phosphorylated at T606 by its association partner cyclin G-associated kinase (GAK). This phosphorylation was required for proper cell proliferation and tumor growth of cells implanted into nude mice. Immunofluorescence analysis showed that the localization of CHC-pT606 signals changed during mitosis. CHC-pT606 signals localized in the nucleus and at the centrosome during interphase, whereas CHC signals were mostly cytoplasmic. Co-immunoprecipitation suggested that CHC formed a complex with GAK and polo-like kinase 1 (PLK1). Depletion of GAK using siRNA induced metaphase arrest and aberrant localization of CHC-pT606, which abolished Kiz-pT379 (as a phosphorylation target of PLK1) signals on chromatin at metaphase. Taken together, we propose that the GAK_CHC-pT606_PLK1_Kiz-pT379 axis plays a role in proliferation of cancer cells.

Keywords: CHC; GAK; Kizuna; PLK1; centrosome; mitosis.

Publication types

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

MeSH terms

Animals
Cell Cycle Proteins / metabolism
Cell Nucleus / metabolism
Centrosome / metabolism
Clathrin Heavy Chains / genetics
Clathrin Heavy Chains / metabolism*
Female
Gene Knockdown Techniques
HeLa Cells
Heterografts
Humans
Interphase / genetics
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism*
Metaphase / genetics*
Mice
Mice, Inbred BALB C
Mice, Nude
Phosphorylation
Polo-Like Kinase 1
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism*
Proto-Oncogene Proteins / metabolism
Spindle Apparatus / metabolism
Transfection
Tumor Burden / genetics

Substances

CLTC protein, human
Cell Cycle Proteins
Intracellular Signaling Peptides and Proteins
Proto-Oncogene Proteins
Clathrin Heavy Chains
GAK protein, human
Protein Serine-Threonine Kinases

Grants and funding

This work was supported in part by Grants-in-Aid for Scientific Research (S) (No. 15101006), Scientific Research (B) (Nos. 20370081 and 23370086), and Exploratory Research (No. 21651085) to H.N. from the Japan Society for the Promotion of Science (JSPS).