Abstract

BackgroundWhite Spot Syndrome Virus, a member of the virus family Nimaviridae, is a large dsDNA virus infecting shrimp and other crustacean species. Although limited information is available on the mode of transcription, previous data suggest that WSSV gene expression occurs in a coordinated and cascaded fashion. To search in silico for conserved promoter motifs (i) the abundance of all 4 through 8 nucleotide motifs in the upstream sequences of WSSV genes relative to the complete genome was determined, and (ii) a MEME search was performed in the upstream sequences of either early or late WSSV genes, as assigned by microarray analysis. Both methods were validated by alignments of empirically determined 5' ends of various WSSV mRNAs.ResultsThe collective information shows that the upstream region of early WSSV genes, containing a TATA box and an initiator, is similar to Drosophila RNA polymerase II core promoter sequences, suggesting utilization of the cellular transcription machinery for generating early transcripts. The alignment of the 5' ends of known well-established late genes, including all major structural protein genes, identified a degenerate motif (ATNAC) which could be involved in WSSV late transcription. For these genes, only one contained a functional TATA box. However, almost half of the WSSV late genes, as previously assigned by microarray analysis, did contain a TATA box in their upstream region.ConclusionThe data may suggest the presence of two separate classes of late WSSV genes, one exploiting the cellular RNA polymerase II system for mRNA synthesis and the other generating messengers by a new virus-induced transcription mechanism.

Highlights

  • White Spot Syndrome Virus, a member of the virus family Nimaviridae, is a large dsDNA virus infecting shrimp and other crustacean species

  • As important promoter motifs are overrepresented in the 5' upstream regions of baculoviruses genes [26], we studied the relative abundance of all 4 through 8 nucleotide motifs in the upstream regions of White Spot Syndrome Virus (WSSV) genes compared to the complete WSSV genomic sequence

  • Promoter analysis using the enumeration method In a search for putative WSSV regulatory promoter elements, we compared the abundance of all 4, 5, 6, 7 or 8 nucleotide motifs in the 100 and 200 nt upstream sequences of all WSSV genes relative to their presence in the complete WSSV genomic sequence. This method will be referred to as the enumeration method in the remaining part of the article. This enumeration method was applied on the genome sequences of the type species of more extensively studied large ds DNA viruses mentioned in the introduction: AcMNPV (Autographa californica Multinucleopolyhedrovirus; Baculovirus), Human Herpes Virus 1 (HHV1; Herpesvirus), Vaccinia

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Summary

Introduction

White Spot Syndrome Virus, a member of the virus family Nimaviridae, is a large dsDNA virus infecting shrimp and other crustacean species. To search in silico for conserved promoter motifs (i) the abundance of all 4 through 8 nucleotide motifs in the upstream sequences of WSSV genes relative to the complete genome was determined, and (ii) a MEME search was performed in the upstream sequences of either early or late WSSV genes, as assigned by microarray analysis Both methods were validated by alignments of empirically determined 5' ends of various WSSV mRNAs. White Spot Syndrome Virus (WSSV), type species of the virus family Nimaviridae (genus whispovirus), is a pathogen of major economic importance in cultured penaeid shrimp [1,2]. WSSV particles have been mainly detected in the nuclei of infected cells, indicating that transcription, replication and virion assembly probably occur in the nucleus [5,6,7,8]. Around 50 major or minor virion protein genes have been identified on the genome [12,13,14,15]

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