The small nonstructural protein NP1 of human bocavirus 1 directly interacts with Ku70 and RPA70 and facilitates viral DNA replication

Kang Ning; Zekun Wang; Fang Cheng; Ziying Yan; Jianming Qiu

doi:10.1371/journal.ppat.1010578

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The small nonstructural protein NP1 of human bocavirus 1 directly interacts with Ku70 and RPA70 and facilitates viral DNA replication

Journal article

Open access

Peer reviewed

The small nonstructural protein NP1 of human bocavirus 1 directly interacts with Ku70 and RPA70 and facilitates viral DNA replication

Kang Ning, Zekun Wang, Fang Cheng, Ziying Yan and Jianming Qiu

PLoS pathogens, Vol.18(6), pp.e1010578-e1010578

06/2022

DOI: 10.1371/journal.ppat.1010578

PMCID: PMC9197078

PMID: 35653410

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Abstract

Human bocavirus 1 (HBoV1), a member of the genus Bocaparvovirus of the family Parvoviridae, causes acute respiratory tract infections in young children. Well-differentiated pseudostratified human airway epithelium cultured at an air-liquid interface (HAE-ALI) is an ideal in vitro culture model to study HBoV1 infection. Unique to other parvoviruses, bocaparvoviruses express a small nonstructured protein NP1 of ~25 kDa from an open reading frame (ORF) in the center of the viral genome. NP1 plays an important role in viral DNA replication and pre-mRNA processing. In this study, we performed an affinity purification assay to identify HBoV1 NP1-inteacting proteins. We identified that Ku70 and RPA70 directly interact with the NP1 at a high binding affinity, characterized with an equilibrium dissociation constant (KD) of 95 nM and 122 nM, respectively. Furthermore, we mapped the key NP1-interacting domains of Ku70 at aa266-439 and of RPA70 at aa181-422. Following a dominant negative strategy, we revealed that the interactions of Ku70 and RPA70 with NP1 play a significant role in HBoV1 DNA replication not only in an in vitro viral DNA replication assay but also in HBoV1-infected HAE-ALI cultures. Collectively, our study revealed a novel mechanism by which HBoV1 NP1 enhances viral DNA replication through its direct interactions with Ku70 and RPA70.

Child

Child, Preschool

DNA Replication

DNA, Viral - genetics

DNA, Viral - metabolism

Genome, Viral

Human bocavirus - genetics

Human bocavirus - metabolism

Humans

Parvoviridae Infections

Virus Replication - genetics

Details

Title: Subtitle: The small nonstructural protein NP1 of human bocavirus 1 directly interacts with Ku70 and RPA70 and facilitates viral DNA replication
Creators: Kang Ning - University of Kansas Medical Center
Zekun Wang - University of Kansas Medical Center
Fang Cheng - University of Kansas Medical Center
Ziying Yan - University of Iowa
Jianming Qiu - University of Kansas Medical Center
Resource Type: Journal article
Publication Details: PLoS pathogens, Vol.18(6), pp.e1010578-e1010578
DOI: 10.1371/journal.ppat.1010578
PMID: 35653410
PMCID: PMC9197078
NLM abbreviation: PLoS Pathog
ISSN: 1553-7374
eISSN: 1553-7374
Publisher: Public Library of Science
Grant note: DOI: 10.13039/100000060, name: National Institute of Allergy and Infectious Diseases, award: AI150877; DOI: 10.13039/100000060, name: National Institute of Allergy and Infectious Diseases, award: AI139572; DOI: 10.13039/100000060, name: National Institute of Allergy and Infectious Diseases, award: AI151542; DOI: 10.13039/100000062, name: National Institute of Diabetes and Digestive and Kidney Diseases, award: DK054759; DOI: 10.13039/100000897, name: Cystic Fibrosis Foundation, award: YAN19XX0
Language: English
Date published: 06/2022
Academic Unit: Anatomy and Cell Biology
Record Identifier: 9984284334002771

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