Splice Junction Map of Simian Parvovirus Transcripts

Abstract

The transcription map of simian parvovirus (SPV), an Erythrovirus similar to Parvovirus B19, was investigated. RNA was extracted from tissues of experimentally infected cynomolgus macaques and subjected to reverse transcription-PCR with SPV-specific primers. The PCR products were cloned and sequenced to identify splice junctions. A total of 14 distinct sequences were identified as putative partial transcripts. Of these, 13 were spliced; a single unspliced transcript putatively encoded NS1. Sequence analysis revealed that spliced partial transcripts may encode portions of open reading frames for the major capsid proteins VP1 and VP2 and smaller, unknown proteins. These unspliced and spliced transcripts and putative proteins encoded by SPV were similar to those of B19. Initial splice junctions at nucleotides 279 and 333 were analogous to those at nucleotides 406 and 441, respectively, in B19. Seven of the 10 splices identified had typical GT/AG donor/ acceptor junctions. The splice sites were confirmed by Northern blotting and autoradiography. In contrast to B19, which has a maximum of two splices per transcript, up to three splices were observed in SPV transcripts. A spliced transcript putatively encoding a truncated version of NS1, as seen with minute virus of mice and adeno-associated virus 2, was also observed. The findings indicate that that the splicing pattern of transcripts of SPV and B19 is similar, but SPV also has coding strategies in common with other parvoviruses.Simian parvovirus (SPV) was first identified in cynomolgus macaques suffering from severe anemia as a consequence of erythroid cell destruction (20). These animals were immunosuppressed due to a concurrent infection with type D simian retrovirus (22). Clinically, this was similar to cases of human infection with parvovirus B19, which is responsible for causing transient aplastic crisis in individuals with underlying hemolysis and red cell aplasia in immunosuppressed individuals, such as AIDS patients (9,22). B19 infection also results in hydrops fetalis and fetal death in pregnant women (9), sequelae also observed with experimental SPV infection in pregnant female cynomolgus macaques (M. G. O'Sullivan, J. C. Veille, W. A. Block, A. R. Turner, N. S. Young, and K. E. Brown, Abstr. 17th Annu. Meet. Am. Soc. Virol., abstr. W22-9, 1998).SPV was classified as a parvovirus considering molecular similarities to other parvoviruses, i.e., adeno-associated virus 2 (prototype dependovirus), the autonomous parvoviruses (the prototype being minute virus of mice), and parvovirus B19 (prototype erythrovirus). In general, members of the subfamily Parvovirinae have a small genome of about 5 kb composed of single-stranded DNA with terminal palindromic sequences that serve as primers for the synthesis of the complementary strand. Only the positive strand of these viruses encodes proteins, suggesting that the positive-sense mRNA is transcribed from the minus-strand genomic DNA (3). To maximize the coding capacity, a number of promoters and all three reading frame...

Biology, Genome Organization, and Evolution of Parvoviruses in Marine Shrimp

“…However, only six of the seven nucleotides at the predicted translation initiation site of right ORF match with the consensus Kozak sequence, indicating the ORF as an "adequately expressed" gene (Kozak, 2002). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004).…”

Biology, Genome Organization, and Evolution of Parvoviruses in Marine Shrimp

“…However, only six of the seven nucleotides at the predicted translation initiation site of right ORF match with the consensus Kozak sequence, indicating the ORF as an "adequately expressed" gene (Kozak, 2002). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004).…”

Comparison of the Transcription Profile of Simian Parvovirus with That of the Human Erythrovirus B19 Reveals a Number of Unique Features

“…3), which reflected unspliced RNA and RNA using a donor at nt 3208, consistent with the RT-PCR and 5Ј RACE and sequencing results described above. This ratio was compatible with the splicing of A3 only to D3, and RT-PCR and sequence analysis did not identify any mRNAs spliced between the D1 or D2 donor sites and the A3 acceptor site (data not shown) (31).…”

Simian Parvovirus Infection: A Potential Zoonosis

“…Sequencing showed that, in both the human and the macaque cultures, the major splice junction was between nt 279 and 1792. Additional splice junctions between nt 279 and 1740 and nt 333 and 1792, as observed in SPV fetal infection [15], were also observed in SPV infection of human bone marrow mononuclear cells.…”

Biology, Genome Organization, and Evolution of Parvoviruses in Marine Shrimp

“…However, only six of the seven nucleotides at the predicted translation initiation site of right ORF match with the consensus Kozak sequence, indicating the ORF as an "adequately expressed" gene (Kozak, 2002). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004).…”

Comparison of the Transcription Profile of Simian Parvovirus with That of the Human Erythrovirus B19 Reveals a Number of Unique Features

“…3), which reflected unspliced RNA and RNA using a donor at nt 3208, consistent with the RT-PCR and 5Ј RACE and sequencing results described above. This ratio was compatible with the splicing of A3 only to D3, and RT-PCR and sequence analysis did not identify any mRNAs spliced between the D1 or D2 donor sites and the A3 acceptor site (data not shown) (31).…”

Simian Parvovirus Infection: A Potential Zoonosis

“…Sequencing showed that, in both the human and the macaque cultures, the major splice junction was between nt 279 and 1792. Additional splice junctions between nt 279 and 1740 and nt 333 and 1792, as observed in SPV fetal infection [15], were also observed in SPV infection of human bone marrow mononuclear cells.…”

Biology, Genome Organization, and Evolution of Parvoviruses in Marine Shrimp

“…However, only six of the seven nucleotides at the predicted translation initiation site of right ORF match with the consensus Kozak sequence, indicating the ORF as an "adequately expressed" gene (Kozak, 2002). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004). Similar observations have been made in HPV of P. monodon (Sukhumsirichart et al, 2006), an iteravirusinfecting Dendrolimus punctatus (Wang et al, 2005), a human pathogenic B19 parvovirus (Ozawa et al, 1987) and a simian parvovirus (Vashisht, Faaberg, Aber, Brown, & O'Sullivan, 2004).…”

Comparison of the Transcription Profile of Simian Parvovirus with That of the Human Erythrovirus B19 Reveals a Number of Unique Features

“…3), which reflected unspliced RNA and RNA using a donor at nt 3208, consistent with the RT-PCR and 5Ј RACE and sequencing results described above. This ratio was compatible with the splicing of A3 only to D3, and RT-PCR and sequence analysis did not identify any mRNAs spliced between the D1 or D2 donor sites and the A3 acceptor site (data not shown) (31).…”

Simian Parvovirus Infection: A Potential Zoonosis

“…Sequencing showed that, in both the human and the macaque cultures, the major splice junction was between nt 279 and 1792. Additional splice junctions between nt 279 and 1740 and nt 333 and 1792, as observed in SPV fetal infection [15], were also observed in SPV infection of human bone marrow mononuclear cells.…”