Structure-function insights into the dual role of African swine fever virus pB318L: A typical geranylgeranyl-diphosphate synthase and a nuclear import protein

Hai-Fan Zhao; Ying Wang; Xiao-Hong Liu; Xian-Hui Liu; Zhi Geng; Zeng-Qiang Gao; Li Huang; Chang-Jiang Weng; Yu-Hui Dong; Heng Zhang

doi:10.1016/j.virs.2025.03.013

April 2025

Citation: Hai-Fan Zhao, Ying Wang, Xiao-Hong Liu, Xian-Hui Liu, Zhi Geng, Zeng-Qiang Gao, Li Huang, Chang-Jiang Weng, Yu-Hui Dong, Heng Zhang. Structure-function insights into the dual role of African swine fever virus pB318L: A typical geranylgeranyl-diphosphate synthase and a nuclear import protein .VIROLOGICA SINICA, 2025, 40(2) : 236-246. http://dx.doi.org/10.1016/j.virs.2025.03.013

Structure-function insights into the dual role of African swine fever virus pB318L: A typical geranylgeranyl-diphosphate synthase and a nuclear import protein

Hai-Fan Zhao ^a,b ,
Ying Wang ^b,c ,
Xiao-Hong Liu ^d ,
Xian-Hui Liu ^e ,
Zhi Geng ^b ,
Zeng-Qiang Gao ^b ,
Li Huang ^d ,
Chang-Jiang Weng ^d ,
Yu-Hui Dong ^{b
,,} ,
Heng Zhang ^{b
,,}

a. School of Life Sciences, University of Science and Technology of China, Hefei 230027, China;
b. Beijing Synchrotron Radiation Facility, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049, China;
c. Key Laboratory of Integrated Pest Management on Crops in East China, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China;
d. State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150040, China;
e. College of Animal Science and National Engineering Center for Swine Breeding Industry, South China Agricultural University, Guangzhou 510642, China

Corresponding author: Yu-Hui Dong, dongyh@ihep.ac.cn
Heng Zhang, zhangheng@ihep.ac.cn
Received Date: 24 July 2024
Accepted Date: 27 March 2025
Available online: 02 April 2025

Abstract

African swine fever virus (ASFV) pB318L is an important protein for viral replication that acts as a membrane-bound trans-geranylgeranyl-diphosphate synthase (GGPPS) catalyzing the condensation of isopentenyl diphosphate (IPP) with allylic diphosphates. Recently we solved the crystal structure pB318L lacking N-terminal transmembrane region and performed a preliminary structural analysis. In this study, structure-based mutagenesis study and geranylgeranyl pyrophosphate (GGPP) production assay further revealed the key residues for the GGPPS activity. Structural comparison showed pB318L displays a strong similarity to typical GGPPSs instead of protein prenyltransferases. The phylogenetic analysis indicated pB318L may share a common ancestor with the GGPPSs from Brassicaceae plants rather than from its natural host. The subcellular localization analysis showed pB318L is localized in both nucleus and cytoplasm (including the endoplasmic reticulum membrane and mitochondria outer membrane). A unique N-terminal nuclear localization signal (NLS) following the transmembrane region was discovered in pB318L and the NLS was confirmed to be required for the nuclear import. We further revealed the NLS plays an essential role in the interaction with nuclear transporter karyopherin subunit alpha 1 (KPNA1). Their interaction may suppress signal transducers and activators of transcription 1 (STAT1) translocation and subsequently competitively inhibit nuclear import of IFN-stimulated gene factor 3 (ISGF3) complex. Our biochemical, structural and cellular analyses provide novel insights to pB318L that acts as an essential GGPPS that promotes viral replication and as a nuclear import protein that may be involved in immune evasion of ASFV.
- African swine fever virus (ASFV)
- , Prenyltransferase
- , Geranylgeranyl-diphosphate synthase
- , Crystal structure
- , Nuclear localization signal
- , Immune evasion

Electronic Supplementary Material

10.1016j.virs.2025.03.013-ESM.docx
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