Molecular Characterization of Infectious Clones of the Minute Virus of Canines Reveals Unique Features of Bocaviruses

Sun, Yuning; Chen, Aaron Yun; Cheng, Fang; Guan, Wuxiang; Johnson, F. Brent; Qiu, Jianming

doi:10.1128/jvi.02569-08

Cited by 135 publications

(231 citation statements)

References 66 publications

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“…BPV is the prototype virus of this genus. The genome consists of ssDNA that is composed of about 5515 nt (Qiu et al, 2007;Sun et al, 2009). The genome has three ORFs: the left ORF encodes the non-structural proteins NS1 and NS2, the central ORF encodes the non-structural protein NP-1, and the right ORF encodes two or three structural proteins VP1, VP2 and VP3 (Johnson & Hoggan, 1973;Qiu et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Bovine parvovirus uses clathrin-mediated endocytosis for cell entry

Dudleenamjil¹,

Lin²,

Dredge³

et al. 2010

Journal of General Virology

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Entry events of bovine parvovirus (BPV) were studied. Transmission electron micrographs of infected cells showed virus particles in cytoplasmic vesicles. Chemical inhibitors that block certain aspects of the cellular machinery were employed to assess viral dependency upon those cellular processes. Chlorpromazine, ammonium chloride, chloroquine and bafilamicin A1 were used to inhibit acidification of endosomes and clathrin-associated endocytosis. Nystatin was used as an inhibitor of the caveolae pathway. Cytochalasin D and ML-7 were used to inhibit actin and myosin functions, respectively. Nocodazole and colchicine were employed to inhibit microtubule activity. Virus entry was assessed by measuring viral transcription using real-time PCR, synthesis of capsid protein and assembly of infectious progeny virus in the presence of inhibitor blockage. The results indicated that BPV entry into embryonic bovine trachael cells utilizes endocytosis in clathrin-coated vesicles, is dependent upon acidification, and appears to be associated with actin and microtubule dependency. Evidence for viral entry through caveolae was not obtained. These findings provide a fuller understanding of the early cell-entry events of the replication cycle for members of the genus Bocavirus.

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Section: Introductionmentioning

confidence: 99%

Bovine parvovirus uses clathrin-mediated endocytosis for cell entry

Dudleenamjil¹,

Lin²,

Dredge³

et al. 2010

Journal of General Virology

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“…Like members in the genus Bocavirus (8,9) and Aleutian mink disease virus (10), also of the Parvoviridae family, the transcription profile of B19V is unusual for a DNA virus in that all of the mRNA transcripts are generated from a single precursor mRNA (pre-mRNA) transcribed from a single promoter at map unit 6 (P6) and feature alternative polyadenylation and splicing (see Fig. 1A) (11,12).…”

Section: Human Parvovirus B19 (B19v)mentioning

confidence: 99%

Internal Polyadenylation of the Parvovirus B19 Precursor mRNA Is Regulated by Alternative Splicing

Guan

Huang

Cheng

et al. 2011

Journal of Biological Chemistry

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Alternative processing of parvovirus B19 (B19V) pre-mRNA is critical to generating appropriate levels of B19V mRNA transcripts encoding capsid proteins and small nonstructural proteins. Polyadenylation of the B19V pre-mRNA at the proximal polyadenylation site ((pA)p), which prevents generation of fulllength capsid proteins encoding mRNA transcripts, has been suggested as a step that blocks B19V permissiveness. We report here that efficient splicing of the B19V pre-mRNA within the first intron (upstream of the (pA)p site) stimulated the polyadenylation; in contrast, splicing of the B19V pre-mRNA within the second intron (in which the (pA)p site resides) interfered with the polyadenylation, leading to the generation of a sufficient number of B19V mRNA transcripts polyadenylated at the distal polyadenylation site ((pA)d). We also found that splicing within the second intron and polyadenylation at the (pA)p site compete during processing of the B19V pre-mRNA. Furthermore, we discovered that the U1 RNA that binds to the 5 splice donor site of the second intron is fully responsible for inhibiting polyadenylation at the (pA)p site, whereas actual splicing, and perhaps assembly of the functional spliceosome, is not required. Finally, we demonstrated that inhibition of B19V pre-mRNA splicing within the second intron by targeting an intronic splicing enhancer using a Morpholino antisense oligonucleotide prevented B19V mRNA transcripts polyadenylated at the (pA)d site during B19V infection of human erythroid progenitors. Thus, our study reveals the mechanism by which alternative splicing coordinates alternative polyadenylation to generate full-length B19V mRNA transcripts at levels sufficient to support productive B19V infection. Human parvovirus B19 (B19V)2 is a member of the genus Erythrovirus of the family Parvoviridae. B19V causes several diseases in humans (1), the most common of which is erythema infectiosum or Fifth disease. Other diseases include acute and chronic arthropathy, transient aplastic crisis (infection of patients with a high rate of red blood cell turnover), pure red cell aplasia (infection of immunocompromised patients), and hydrops fetalis (infection of pregnant women). Like other parvoviruses, B19V contains a linear single-stranded DNA genome of approximately (ϳ)5.6 kb, which is encapsidated by an icosahedrally symmetric capsid without an envelope (2, 3). In addition to infecting only humans, B19V infection shows a remarkable tropism for human erythroid progenitor cells (4 -7).Like members in the genus Bocavirus (8, 9) and Aleutian mink disease virus (10), also of the Parvoviridae family, the transcription profile of B19V is unusual for a DNA virus in that all of the mRNA transcripts are generated from a single precursor mRNA (pre-mRNA) transcribed from a single promoter at map unit 6 (P6) and feature alternative polyadenylation and splicing (see Fig. 1A) (11, 12). The only unspliced mRNA encoding the large nonstructural protein (NS1) is polyadenylated at the proximal poly(A) site ((pA)p) and retains...

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“…Thus, genome replication is a necessary precursor to transcription. The genomic organization of HBoV, the second human-pathogenic parvovirus known (discovered after Parvovirus B19), closely resembles the other known bocaviruses BPV and CnMV (Allander et al, 2005;Kaplan et al, 2006;Ma et al, 2006;Sun et al, 2009). The mid-ORF product of HBoV is homologous to NP-1 of BPV and CnMV, and these proteins have 47% amino acid identity.…”

Section: Introductionmentioning

confidence: 99%

“…It is a contamination risk for commercially prepared bovine serum and bovine-derived products. The genome consists of single stranded DNA that is composed of about 5515 nucleotides (Chen et al, 1986;Qiu et al, 2007;Sun, et al, 2009). The genome contains non-identical palindromic sequences at the two ends.…”

Section: Introductionmentioning

confidence: 99%