Endosomal phosphatidylserine is critical for the YAP signalling pathway in proliferating cells

Tatsuyuki Matsudaira; Kojiro Mukai; Taishin Noguchi; Junya Hasegawa; Tomohisa Hatta; Shun-Ichiro Iemura; Tohru Natsume; Norio Miyamura; Hiroshi Nishina; Jun Nakayama; Kentaro Semba; Takuya Tomita; Shigeo Murata; Hiroyuki Arai; Tomohiko Taguchi

doi:10.1038/s41467-017-01255-3

Endosomal phosphatidylserine is critical for the YAP signalling pathway in proliferating cells

Nat Commun. 2017 Nov 1;8(1):1246. doi: 10.1038/s41467-017-01255-3.

Authors

Tatsuyuki Matsudaira¹, Kojiro Mukai¹, Taishin Noguchi¹, Junya Hasegawa¹, Tomohisa Hatta², Shun-Ichiro Iemura³, Tohru Natsume², Norio Miyamura⁴, Hiroshi Nishina⁴, Jun Nakayama⁵, Kentaro Semba⁵, Takuya Tomita⁶, Shigeo Murata⁶, Hiroyuki Arai^{7

8

9}, Tomohiko Taguchi^{10

11}

Affiliations

¹ Department of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
² Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology, 2-3-26, Aomi, Koto-ku, Tokyo, 135-0064, Japan.
³ Medical Industry Translational Research Center, Fukushima Medical University, 1, Hikarigaoka, Fukushima, 960-1295, Japan.
⁴ Department of Developmental and Regenerative Biology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
⁵ Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, 2-2, Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan.
⁶ Laboratory of Protein Metabolism, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
⁷ Department of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. harai@mol.f.u-tokyo.ac.jp.
⁸ Pathological Cell Biology Laboratory, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. harai@mol.f.u-tokyo.ac.jp.
⁹ AMED-CREST, Japan Agency for Medical Research and Development, 1-7-1, Otemachi, Chiyoda-ku, Tokyo, 100-0004, Japan. harai@mol.f.u-tokyo.ac.jp.
¹⁰ Department of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. tom_taguchi@mol.f.u-tokyo.ac.jp.
¹¹ Pathological Cell Biology Laboratory, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. tom_taguchi@mol.f.u-tokyo.ac.jp.

Abstract

Yes-associated protein (YAP) is a recently discovered growth-promoting transcription coactivator that has been shown to regulate the malignancy of various cancers. How YAP is regulated is not fully understood. Here, we show that one of the factors regulating YAP is phosphatidylserine (PS) in recycling endosomes (REs). We use proximity biotinylation to find proteins proximal to PS. Among these proteins are YAP and multiple proteins related to YAP signalling. Knockdown of ATP8A1 (an RE PS-flippase) or evectin-2 (an RE-resident protein) and masking of PS in the cytoplasmic leaflet of membranes, all suppress nuclear localization of YAP and YAP-dependent transcription. ATP8A1 knockdown increases the phosphorylated (activated) form of Lats1 that phosphorylates and inactivates YAP, whereas evectin-2 knockdown reduces the ubiquitination and increased the level of Lats1. The proliferation of YAP-dependent metastatic cancer cells is suppressed by knockdown of ATP8A1 or evectin-2. These results suggest a link between a membrane phospholipid and cell proliferation.

Publication types

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

MeSH terms

Adaptor Proteins, Signal Transducing / metabolism*
Adenosine Triphosphatases / genetics*
Animals
Biotinylation
COS Cells
Cell Nucleus / metabolism
Cell Proliferation
Chlorocebus aethiops
Endosomes / metabolism
Gene Knockdown Techniques
HEK293 Cells
Humans
Membrane Proteins / genetics*
Phosphatidylserines / metabolism*
Phospholipid Transfer Proteins / genetics*
Phosphoproteins / metabolism*
Phosphorylation
Protein Serine-Threonine Kinases / metabolism*
Protein Transport / genetics
Signal Transduction
Transcription Factors
Ubiquitination
YAP-Signaling Proteins

Substances

Adaptor Proteins, Signal Transducing
Membrane Proteins
PLEKHB2 protein, human
Phosphatidylserines
Phospholipid Transfer Proteins
Phosphoproteins
Transcription Factors
YAP-Signaling Proteins
YAP1 protein, human
LATS1 protein, human
Protein Serine-Threonine Kinases
ATP8A1 protein, human
Adenosine Triphosphatases