P300-mediated NEDD4 acetylation drives ebolavirus VP40 egress by enhancing NEDD4 ligase activity.

Linliang Zhang,Dongning Jin,Mingzhou Chen,Yi Yang,Lei Yin,Majuan Chen,Shixiong Zhou,Linjuan Wu,Jie Yan,Yali Qin,Amy L Hartman

doi:10.1371/journal.ppat.1009616

Abstract

The final stage of Ebola virus (EBOV) replication is budding from host cells, where the matrix protein VP40 is essential for driving this process. Many post-translational modifications such as ubiquitination are involved in VP40 egress, but acetylation has not been studied yet. Here, we characterize NEDD4 is acetylated at a conserved Lys667 mediated by the acetyltransferase P300 which drives VP40 egress process. Importantly, P300-mediated NEDD4 acetylation promotes NEDD4-VP40 interaction which enhances NEDD4 E3 ligase activity and is essential for the activation of VP40 ubiquitination and subsequent egress. Finally, we find that Zaire ebolavirus production is dramatically reduced in P300 knockout cell lines, suggesting that P300-mediated NEDD4 acetylation may have a physiological effect on Ebola virus life cycle. Thus, our study identifies an acetylation-dependent regulatory mechanism that governs VP40 ubiquitination and provides insights into how acetylation controls EBOV VP40 egress.

Highlights

Ebola virus (EBOV) is one of the deadliest pathogens, causing fatal hemorrhagic fever diseases in humans and primates [1,2]
We find that P300-mediated NEDD4 acetylation facilitates EBOV egress
We found that when all proteins were detected at equivalent levels in cell lysates, the HDAC deacetylase family inhibitors trichostatin A (TSA) (Fig 1A, lane 2) and suberoylanilide hydroxamic acid (SAHA) (Fig 1A, lane 3) could facilitate the release of VP40 VLPs, while the increased levels of acetyl-tubulin implied the two inhibitors had worked (Fig 1A)

Summary

Introduction

Ebola virus (EBOV) is one of the deadliest pathogens, causing fatal hemorrhagic fever diseases in humans and primates [1,2]. The outbreak in the Democratic Republic of Congo during June 2020 caused significant alarm. The core nucleocapsid structure of EBOV is composed of viral RNA, nucleoprotein (NP), viral protein (VP) 35, VP30, and the polymerase L [3,4,5]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

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