A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium

Human bocavirus 1 (HBoV1) infects well-differentiated (polarized) human airway epithelium (HAE) cultured at an air-liquid interface (ALI). In the present study, we applied next-generation RNA sequencing to investigate the genome-wide transcription profile of HBoV1, including viral mRNA and small RNA...

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Main Authors: Wei Zou, Min Xiong, Xuefeng Deng, John F. Engelhardt, Ziying Yan, Jianming Qiu
Format: Article
Language:English
Published: MDPI AG 2019-01-01
Series:Viruses
Subjects:
Online Access:http://www.mdpi.com/1999-4915/11/1/33
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spelling doaj-adbb13e9127d47a1a835885fdbe7ad102020-11-25T00:16:51ZengMDPI AGViruses1999-49152019-01-011113310.3390/v11010033v11010033A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway EpitheliumWei Zou0Min Xiong1Xuefeng Deng2John F. Engelhardt3Ziying Yan4Jianming Qiu5Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USAThe Children’s Mercy Hospital, School of Medicine, University of Missouri Kansas City, Kansas City, MO 64108, USADepartment of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USADepartment of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USADepartment of Anatomy and Cell Biology, University of Iowa, Iowa City, IA 52242, USADepartment of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USAHuman bocavirus 1 (HBoV1) infects well-differentiated (polarized) human airway epithelium (HAE) cultured at an air-liquid interface (ALI). In the present study, we applied next-generation RNA sequencing to investigate the genome-wide transcription profile of HBoV1, including viral mRNA and small RNA transcripts, in HBoV1-infected HAE cells. We identified novel transcription start and termination sites and confirmed the previously identified splicing events. Importantly, an additional proximal polyadenylation site (pA)p2 and a new distal polyadenylation site (pA)dREH lying on the right-hand hairpin (REH) of the HBoV1 genome were identified in processing viral pre-mRNA. Of note, all viral nonstructural proteins-encoding mRNA transcripts use both the proximal polyadenylation sites [(pA)p1 and (pA)p2] and distal polyadenylation sites [(pA)d1 and (pA)dREH] for termination. However, capsid proteins-encoding transcripts only use the distal polyadenylation sites. While the (pA)p1 and (pA)p2 sites were utilized at roughly equal efficiency for proximal polyadenylation of HBoV1 mRNA transcripts, the (pA)d1 site was more preferred for distal polyadenylation. Additionally, small RNA-seq analysis confirmed there is only one viral noncoding RNA (BocaSR) transcribed from nt 5199–5340 of the HBoV1 genome. Thus, our study provides a systematic and unbiased transcription profile, including both mRNA and small RNA transcripts, of HBoV1 in HBoV1-infected HAE-ALI cultures.http://www.mdpi.com/1999-4915/11/1/33parvovirushuman bocavirus 1RNA-seqtranscription profilehuman airway epithelia
collection DOAJ
language English
format Article
sources DOAJ
author Wei Zou
Min Xiong
Xuefeng Deng
John F. Engelhardt
Ziying Yan
Jianming Qiu
spellingShingle Wei Zou
Min Xiong
Xuefeng Deng
John F. Engelhardt
Ziying Yan
Jianming Qiu
A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
Viruses
parvovirus
human bocavirus 1
RNA-seq
transcription profile
human airway epithelia
author_facet Wei Zou
Min Xiong
Xuefeng Deng
John F. Engelhardt
Ziying Yan
Jianming Qiu
author_sort Wei Zou
title A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
title_short A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
title_full A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
title_fullStr A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
title_full_unstemmed A Comprehensive RNA-seq Analysis of Human Bocavirus 1 Transcripts in Infected Human Airway Epithelium
title_sort comprehensive rna-seq analysis of human bocavirus 1 transcripts in infected human airway epithelium
publisher MDPI AG
series Viruses
issn 1999-4915
publishDate 2019-01-01
description Human bocavirus 1 (HBoV1) infects well-differentiated (polarized) human airway epithelium (HAE) cultured at an air-liquid interface (ALI). In the present study, we applied next-generation RNA sequencing to investigate the genome-wide transcription profile of HBoV1, including viral mRNA and small RNA transcripts, in HBoV1-infected HAE cells. We identified novel transcription start and termination sites and confirmed the previously identified splicing events. Importantly, an additional proximal polyadenylation site (pA)p2 and a new distal polyadenylation site (pA)dREH lying on the right-hand hairpin (REH) of the HBoV1 genome were identified in processing viral pre-mRNA. Of note, all viral nonstructural proteins-encoding mRNA transcripts use both the proximal polyadenylation sites [(pA)p1 and (pA)p2] and distal polyadenylation sites [(pA)d1 and (pA)dREH] for termination. However, capsid proteins-encoding transcripts only use the distal polyadenylation sites. While the (pA)p1 and (pA)p2 sites were utilized at roughly equal efficiency for proximal polyadenylation of HBoV1 mRNA transcripts, the (pA)d1 site was more preferred for distal polyadenylation. Additionally, small RNA-seq analysis confirmed there is only one viral noncoding RNA (BocaSR) transcribed from nt 5199–5340 of the HBoV1 genome. Thus, our study provides a systematic and unbiased transcription profile, including both mRNA and small RNA transcripts, of HBoV1 in HBoV1-infected HAE-ALI cultures.
topic parvovirus
human bocavirus 1
RNA-seq
transcription profile
human airway epithelia
url http://www.mdpi.com/1999-4915/11/1/33
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