title: Parvovirus Expresses a Small Noncoding RNA That Plays an Essential Role in Virus Replication.
abstract: Human bocavirus 1 (HBoV1) belongs to the speciesPrimate bocaparvovirusof the genusBocaparvovirusof theParvoviridaefamily. HBoV1 causes acute respiratory tract infections in young children and has a selective tropism for the apical surface of well-differentiated human airway epithelia (HAE). In this study, we identified an additional HBoV1 gene, bocavirus-transcribed small noncoding RNA (BocaSR), within the 3' noncoding region (nucleotides [nt] 5199 to 5338) of the viral genome of positive sense. BocaSR is transcribed by RNA polymerase III (Pol III) from an intragenic promoter at levels similar to that of the capsid protein-coding mRNA and is essential for replication of the viral DNA in both transfected HEK293 and infected HAE cells. Mechanistically, we showed that BocaSR regulates the expression of HBoV1-encoded nonstructural proteins NS1, NS2, NS3, and NP1 but not NS4. BocaSR is similar to the adenovirus-associated type I (VAI) RNA in terms of both nucleotide sequence and secondary structure but differs from it in that its regulation of viral protein expression is independent of RNA-activated protein kinase (PKR) regulation. Notably, BocaSR accumulates in the viral DNA replication centers within the nucleus and likely plays a direct role in replication of the viral DNA. Our findings reveal BocaSR to be a novel viral noncoding RNA that coordinates the expression of viral proteins and regulates replication of viral DNA within the nucleus. Thus, BocaSR may be a target for antiviral therapies for HBoV and may also have utility in the production of recombinant HBoV vectors.IMPORTANCEHuman bocavirus 1 (HBoV1) is pathogenic to humans, causing acute respiratory tract infections in young children. In this study, we identified a novel HBoV1 gene that lies in the 3' noncoding region of the viral positive-sense genome and is transcribed by RNA polymerase III into a noncoding RNA of 140 nt. This bocavirus-transcribed small RNA (BocaSR) diverges from both adenovirus-associated (VA) RNAs and Epstein-Barr virus-encoded small RNAs (EBERs) with respect to RNA sequence, representing a third species of this kind of Pol III-dependent viral noncoding RNA and the first noncoding RNA identified in autonomous parvoviruses. Unlike the VA RNAs, BocaSR localizes to the viral DNA replication centers of the nucleus and is essential for expression of viral nonstructural proteins independent of RNA-activated protein kinase R and replication of HBoV1 genomes. The identification of BocaSR and its role in virus DNA replication reveals potential avenues for developing antiviral therapies.
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.
authors.forename: Zekun
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.
authors.forename: Weiran
authors.initials: W
authors.lastname: Shen
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Cheng
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Deng
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: John F
authors.initials: JF
authors.lastname: Engelhardt
authors.affiliation: Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Ziying
authors.initials: Z
authors.lastname: Yan
authors.affiliation: Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas, USA jqiu@kumc.edu.
authors.forename: Jianming
authors.initials: J
authors.lastname: Qiu
mesh_terms: D002478:Cells, Cultured; D004261:DNA Replication; D015967:Gene Expression Regulation, Viral; D056661:Human bocavirus / Q000502:physiology*; D006801:Humans; D058727:RNA, Small Untranslated / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D058727:RNA, Small Untranslated
keywords: DNA replication; bocavirus; noncoding RNA; parvovirus