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

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

doi:10.1038/s41467-017-01255-3

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.

Highlights

Yes-associated protein (YAP) is a recently discovered growth-promoting transcription coactivator that has been shown to regulate the malignancy of various cancers
COS-1 cells were stably expressed with a construct consisting of 2xPH, BirA*, and GFP (Fig. 1a)
The increase of phosphorylated YAP was reduced by depletion of Lats1/2 (Supplementary Fig. 3). These results indicated that the concentrated PS in the cytoplasmic leaflet of recycling endosomes (REs) membranes is critical for the nuclear localization of YAP and the transcription of YAP-downstream genes by suppressing Lats[1] activity

Summary

Introduction

Yes-associated protein (YAP) is a recently discovered growth-promoting transcription coactivator that has been shown to regulate the malignancy of various cancers.

Results

Conclusion