Recent Advances in Molecular Biology of Human Bocavirus 1 and Its Applications

Shao, Liting; Shen, Weiran; Wang, Shengqi; Qiu, Jianming

doi:10.3389/fmicb.2021.696604

Cited by 18 publications

(37 citation statements)

References 118 publications

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“…NS1 contains three functional domains, including the N-terminal DNA-binding/endonuclease domain, a helicase domain in the middle, and the C-terminal transcription activation domain, in which the endonuclease and helicase domains are essential to viral DNA replication ( 28 , 68 , 69 ). We noted that it was difficult to purify the large mass of the NS1 protein (NS1-100) in bacteria; thus, we chose to purify the short isoform of NS1, i.e., the NS1-70 protein, which contains the endonuclease and helicase domains and is sufficient to induce a DDR ( 57 ) for an assessment of its interaction with Ku70.…”

Section: Resultsmentioning

confidence: 99%

The Large Nonstructural Protein (NS1) of Human Bocavirus 1 Directly Interacts with Ku70, Which Plays an Important Role in Virus Replication in Human Airway Epithelia

Shao

Ning

Wang

et al. 2022

J Virol

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Human bocavirus 1 (HBoV1), an autonomous human parvovirus, causes acute respiratory tract infections in young children. HBoV1 infects well-differentiated (polarized) human airway epithelium cultured at an air-liquid interface (HAE-ALI). HBoV1 expresses a large nonstructural protein, NS1, that is essential for viral DNA replication. HBoV1 infection of polarized human airway epithelial cells induces a DNA damage response (DDR) that is critical to viral DNA replication involving DNA repair with error-free Y-family DNA polymerases. HBoV1 NS1 or the isoform NS1-70 per se induces a DDR. In this study, using the second-generation proximity-dependent biotin identification (BioID2) approach, we identified that Ku70 is associated with the NS1-BioID2 pulldown complex through a direct interaction with NS1. Bio-layer Interferometry (BLI) assay determined a high binding affinity of the NS1 with Ku70, which has an equilibrium dissociation constant (K D ) value of 0.16 μM and processes the strongest interaction at the C-terminal domain. The association of Ku70 with NS1 was also revealed during HBoV1 infection of HAE-ALI. Knockdown of Ku70 and overexpression of the C-terminal domain of Ku70 significantly decreased HBoV1 replication in HAE-ALI. Thus, our study provides for the first time a direct interaction of a parvovirus large nonstructural protein NS1 with Ku70. IMPORTANCE Parvovirus infection induces a DNA damage response (DDR) that plays a pivotal role in viral DNA replication. The DDR includes activation of ATM (Ataxia telangiectasia mutated), ATR (ATM- and RAD3-related), and DNA-PKcs (DNA-dependent protein kinase catalytic subunit). The large nonstructural protein (NS1) often plays a role in the induction of DDR; however, how the DDR is induced during parvovirus infection or simply by the NS1 is not well studied. Activation of DNA-PKcs has been shown as one of the key DDR pathways in DNA replication of HBoV1. We identified that HBoV1 NS1 directly interacts with Ku70, but not Ku80, of the Ku70/Ku80 heterodimer at a high affinity. This interaction is also important for HBoV1 replication in HAE-ALI. We propose that the interaction of the NS1 with Ku70 recruits the Ku70/Ku80 complex to the viral DNA replication center, which activates DNA-PKcs and facilitates viral DNA replication.

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Section: Resultsmentioning

confidence: 99%

The Large Nonstructural Protein (NS1) of Human Bocavirus 1 Directly Interacts with Ku70, Which Plays an Important Role in Virus Replication in Human Airway Epithelia

Shao

Ning

Wang

et al. 2022

J Virol

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“…Of the five substitutions in NS1 protein found in our samples, two are located in the middle helicase domain [ 9 ] but fall outside of four conserved Walker motifs which execute 3′–5′ helicase function [ 9 , 40 ]. Three substitutions are located in the C-terminal part of NS1 protein, which is predicted to have transcription transactivation capability, but has not been studied [ 9 , 41 ].…”

Section: Discussionmentioning

confidence: 99%

“…HBoV possess a linear, single-stranded DNA genome of 5543 nucleotides (nt) long with non-identical terminal hairpins of 140 and 122 nt that play a key role in virus replication [ 7 , 8 ]. Recent advances in molecular biology research of HBoV1 revealed that during its replication in the polarized/nondividing airway epithelial cells HboV1 expresses six nonstructural proteins: NP1, NS1, NS1-70, NS2, NS3, and NS4, depending on splicing mRNAs within ORF 1 [ 9 ]. The mRNA spliced at the D2-A2 sites results in a shift of the NS1 ORF at the C-terminus and encodes NS1.…”

Section: Introductionmentioning

confidence: 99%

“…The mRNA spliced at the D2-A2 sites results in a shift of the NS1 ORF at the C-terminus and encodes NS1. Unspliced mRNA that reads the D2-A2 intron encodes the NS1-70 protein, and alternative splicing within the NS1-coding region generates mRNAs that encode NS2, NS3 and NS4 [ 9 ]. Moreover, HBoV1 also expresses viral non-coding RNA (BocaSR) and three structural proteins VP1, VP2, and VP3.…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, HBoV1 also expresses viral non-coding RNA (BocaSR) and three structural proteins VP1, VP2, and VP3. The BocaSR is the first identified RNA polymerase III (Pol III) transcribed viral non-coding RNA in small DNA viruses [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

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Prevalence and Molecular Characterization of Human Bocavirus Detected in Croatian Children with Respiratory Infection

Ljubin‐Sternak

Slović

Mijač

et al. 2021

Viruses

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Human bocavirus (HBoV) 1 is considered an important respiratory pathogen, while the role of HBoV2-4 in clinical disease remains somewhat controversial. Since, they are characterized by a rapid evolution, worldwide surveillance of HBoVs’ genetics is necessary. This study explored the prevalence of HBoV genotypes in pediatric patients with respiratory tract infection in Croatia and studied their phylogeny. Using multiplex PCR for 15 respiratory viruses, we investigated 957 respiratory samples of children up to 18 years of age with respiratory tract infection obtained from May 2017 to March 2021 at two different hospitals in Croatia. Amplification of HBoV near-complete genome or three overlapping fragments was performed, sequenced, and their phylogenetic inferences constructed. HBoV was detected in 7.6% children with a median age of 1.36 years. Co-infection was observed in 82.2% samples. Sequencing was successfully performed on 29 HBoV positive samples, and all belonged to HBoV1. Croatian HBoV1 sequences are closely related to strains isolated worldwide, and no phylogenetic grouping based on mono- or co-infection cases or year of isolation was observed. Calculated rates of evolution for HBoV1 were 10−4 and 10−5 substitutions per site and year. Recombination was not detected among sequences from this study.

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Optimization strategies and advances in the research and development of AAV‐based gene therapy to deliver large transgenes

Kolesnik,

Nurtdinov,

Oloruntimehin

et al. 2024

Clinical & Translational Med

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Adeno‐associated virus (AAV)‐based therapies are recognized as one of the most potent next‐generation treatments for inherited and genetic diseases. However, several biological and technological aspects of AAV vectors remain a critical issue for their widespread clinical application. Among them, the limited capacity of the AAV genome significantly hinders the development of AAV‐based gene therapy. In this context, genetically modified transgenes compatible with AAV are opening up new opportunities for unlimited gene therapies for many genetic disorders. Recent advances in de novo protein design and remodelling are paving the way for new, more efficient and targeted gene therapeutics. Using computational and genetic tools, AAV expression cassette and transgenic DNA can be split, miniaturized, shuffled or created from scratch to mediate efficient gene transfer into targeted cells. In this review, we highlight recent advances in AAV‐based gene therapy with a focus on its use in translational research. We summarize recent research and development in gene therapy, with an emphasis on large transgenes (>4.8 kb) and optimizing strategies applied by biomedical companies in the research pipeline. We critically discuss the prospects for AAV‐based treatment and some emerging challenges. We anticipate that the continued development of novel computational tools will lead to rapid advances in basic gene therapy research and translational studies.

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Recent Advances in Molecular Biology of Human Bocavirus 1 and Its Applications

Cited by 18 publications

References 118 publications

The Large Nonstructural Protein (NS1) of Human Bocavirus 1 Directly Interacts with Ku70, Which Plays an Important Role in Virus Replication in Human Airway Epithelia

The Large Nonstructural Protein (NS1) of Human Bocavirus 1 Directly Interacts with Ku70, Which Plays an Important Role in Virus Replication in Human Airway Epithelia

Prevalence and Molecular Characterization of Human Bocavirus Detected in Croatian Children with Respiratory Infection

Optimization strategies and advances in the research and development of AAV‐based gene therapy to deliver large transgenes

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