The Transcription Profile of the Bocavirus Bovine Parvovirus Is Unlike Those of Previously Characterized Parvoviruses

Abstract

The Bocavirus bovine parvovirus generated a single pre-mRNA from a promoter at its left-hand end; however, the pattern of its alternative polyadenylation and splicing was different from that of other parvoviruses. A large left-hand-end open reading frame (ORF) encoded a nonstructural protein of approximately 95 kDa. An abundant, spliced, internally polyadenylated transcript encoded the viral NP1 protein from an ORF in the center of the genome. Transcripts encoding the capsid proteins were polyadenylated in the right-hand terminal palindrome. This is the first published transcription map of a member of the Bocavirus genus of the Parvovirinae.The genus Bocavirus is one of five genera of the subfamily Parvovirinae of the family Parvoviridae (35). The currently recognized members of the Bocavirus genus include bovine parvovirus type 1 (BPV) (1), canine minute virus (32), and the recently identified human bocavirus (HBoV) (4). HBoV, first detected in pooled human samples of lower respiratory tract infections collected by PCR amplification in Sweden (4), has been reported to be associated with acute respiratory illness at incidence rates of between 1.5% and 11.3% worldwide (3, 5, 6, 11-13, 15, 19, 22-24, 34). Isolation of HBoV has not yet been reported.A transcription map of any Bocavirus species based on a nonbiased direct analysis of steady-state RNA has not yet been formally presented. In this report, we describe the transcription map of BPV following infection of permissive bovine turbinate (BT) cells as determined by RNase protection and Northern blotting assays. Like members of the genera Erythrovirus and Amdovirus of the Parvovirinae, the Bocavirus BPV has a single promoter at its left-hand end and uses both an internal and distal polyadenylation site. However, the alternative RNA processing strategy used by BPV generates a transcription profile that is different from that of other characterized parvoviruses.Sequencing of BPV and construction of a nearly full-length BPV clone. Because a previously constructed infectious clone of BPV (33) was no longer available, we recloned the majority of the BPV genome, using a PCR-based strategy, directly from plaque-purified BPV (the prototype Abinanti strain) (1), utilizing the published BPV sequence (GenBank accession no. NC_001540) to design primers. The revised sequence of BPV was deposited in GenBank, and all the nucleotide numbers in the manuscript refer to this revised sequence, unless otherwise specified. This nearly full-length clone, containing the revised BPV sequence (nucleotide [nt] 42 to 5515) in the pBluescript vector (Stratagene), was designated pSK42BPV. This clone was not infectious (data not shown).Our revised BPV sequence showed a number of silent mutations which differed from that of the sequence deposited previously, and these differences may reflect natural variations among isolates. However, in addition, we detected two more significant changes within the NS1 coding region which allowed the opening of the long NS open reading frame 1 (ORF 1) from n...

ELISAs using human bocavirus VP2 virus-like particles for detection of antibodies against HBoV

“…This segment lacks both the left and right-hand palindromic hairpin termini, and therefore, the plasmid containing this sequence of HBoV is not infectious. The genomic organization of HBoV closely resembles that of BPV (Allander et al, 2005;Qiu et al, 2007). There are two major ORFs encoding a nonstructural protein (NS1) and at least the two capsid proteins VP1 and VP2, respectively.…”

Bovine parvovirus uses clathrin-mediated endocytosis for cell entry

“…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).…”

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

“…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).…”