V-ATPase V0a1 promotes Weibel–Palade body biogenesis through the regulation of membrane fission

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Abstract

Membrane fission, the division of a membrane-bound structure into two discrete compartments, is essential for diverse cellular events, such as endocytosis and vesicle/granule biogenesis; however, the process remains unclear. The hemostatic protein von Willebrand factor is produced in vascular endothelial cells and packaged into specialized secretory granules, Weibel-Palade bodies (WPBs) at the trans-Golgi network (TGN). Here, we reported that V0a1, a V-ATPase component, is required for the membrane fission of WPBs. We identified two V0a isoforms in distinct populations of WPBs in cultured endothelial cells, V0a1 and V0a2, on mature and nascent WPBs, respectively. Although WPB buds were formed, WPBs could not separate from the TGN in the absence of V0a1. Screening using dominant-negative forms of known membrane fission regulators revealed protein kinase D (PKD) as an essential factor in biogenesis of WPBs. Further, we showed that the induction of wild-type PKDs in V0a1-depleted cells does not support the segregation of WPBs from the TGN; suggesting a primary role of V0a1 in the membrane fission of WPBs. The identification of V0a1 as a new membrane fission regulator should facilitate the understanding of molecular events that enable membrane fission.

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All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for all blots and graphs shown in the manuscript.

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Yasuo Yamazaki

Department of Molecular Pathogenesis, National Cerebral and Cardiovascular Center, Osaka, Japan

For correspondence
yasuo.yamazaki@ncvc.go.jp

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5297-9837
Yuka Eura

Department of Molecular Pathogenesis, National Cerebral and Cardiovascular Center, Osaka, Japan

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5784-6339
Koichi Kokame

Department of Molecular Pathogenesis, National Cerebral and Cardiovascular Center, Osaka, Japan

For correspondence
kame@ncvc.go.jp

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-9654-6299

Funding

Japan Society for the Promotion of Science (17K07401)

Yasuo Yamazaki

Japan Society for the Promotion of Science (17K07358)

Koichi Kokame

Ministry of Health, Labour and Welfare (JPMH20FC1024)

Koichi Kokame

SENSHIN Medical Research Foundation

Yasuo Yamazaki

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

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This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.