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J Virol
2020 Dec 09;951:. doi: 10.1128/JVI.01594-20.
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Diversity of Sea Star-Associated Densoviruses and Transcribed Endogenous Viral Elements of Densovirus Origin.
Jackson EW
,
Wilhelm RC
,
Johnson MR
,
Lutz HL
,
Danforth I
,
Gaydos JK
,
Hart MW
,
Hewson I
.
Abstract
A viral etiology of sea star wasting syndrome (SSWS) was originally explored with virus-sized material challenge experiments, field surveys, and metagenomics, leading to the conclusion that a densovirus is the predominant DNA virus associated with this syndrome and, thus, the most promising viral candidate pathogen. Single-stranded DNA viruses are, however, highly diverse and pervasive among eukaryotic organisms, which we hypothesize may confound the association between densoviruses and SSWS. To test this hypothesis and assess the association of densoviruses with SSWS, we compiled past metagenomic data with new metagenomic-derived viral genomes from sea stars collected from Antarctica, California, Washington, and Alaska. We used 179 publicly available sea star transcriptomes to complement our approaches for densovirus discovery. Lastly, we focus the study on sea star-associated densovirus (SSaDV), the first sea star densovirus discovered, by documenting its biogeography and putative tissue tropism. Transcriptomes contained only endogenized densovirus elements similar to the NS1 gene, while numerous extant densoviral genomes were recovered from viral metagenomes. SSaDV was associated with nearly all tested species from southern California to Alaska, and in contrast to previous work, we show that SSaDV is one genotype among a high diversity of densoviruses present in sea stars across the West Coast of the United States and globally that are commonly associated with grossly normal (i.e., healthy or asymptomatic) animals. The diversity and ubiquity of these viruses in sea stars confound the original hypothesis that one densovirus is the etiological agent of SSWS.IMPORTANCE The primary interest in sea star densoviruses, specifically SSaDV, has been their association with sea star wasting syndrome (SSWS), a disease that has decimated sea star populations across the West Coast of the United States since 2013. The association of SSaDV with SSWS was originally drawn from metagenomic analysis, which was further studied through field surveys using quantitative PCR (qPCR), with the conclusion that it was the most likely viral candidate in the metagenomic data based on its representation in symptomatic sea stars compared to asymptomatic sea stars. We reexamined the original metagenomic data with additional genomic data sets and found that SSaDV was 1 of 10 densoviruses present in the original data set and was no more represented in symptomatic sea stars than in asymptomatic sea stars. Instead, SSaDV appears to be a widespread, generalist virus that exists among a large diversity of densoviruses present in sea star populations.
FIG 1. Reanalysis of metagenomic data presented by Hewson et al. (11). SSaDV is 1 of 10 densoviruses present in the data set and based on read mapping analysis (≥95% read identity) and is not more abundant in symptomatic than in asymptomatic individuals. (A) Relative abundance of all reads recruited to densovirus genomes. n.s., no significance, based on Welch two-sample t test (P = 0.7697, df = 25.137, t = −0.29592). (B) Read recruitment separated by densovirus genotype.
FIG 2. SSaDV is broadly distributed across the northeastern Pacific Ocean and putatively has a wide tissue tropism. White dots on map indicated PCR-positive samples, and the size of the dot corresponds to total number of PCR-positive samples at each site. Tissue tropism was assessed from 3 sea star species collected from one site (Langley Harbor, WA). The color of each bar corresponds to the anatomical region in the sea star illustration. Prevalence is defined as the number of PCR-positive samples divided by the total number of samples tested for each tissue.
FIG 3. Sea star-associated densoviruses are genetically diverse and are not monophyletic. (A) Cladogram of a maximum likelihood phylogenetic tree of densoviruses based on alignment of amino acid sequences from the NS1 gene. Collapsed nodes represent densovirus genera, while all other branches belong to the genus Ambidensovirus. Red names indicate genomes discovered in this study. White circles represent 90 to 100% bootstrap support. (B) Representative densovirus genome showing genome organization. (C) Histograms of nucleotide and amino acid pairwise identity comparisons between all sea star-associated densoviruses for NS1, NS3, and VP ORFs. Dashed lines indicate mean pairwise identity.
FIG 4. Sea star-associated densoviruses exhibit two genome organizations and contain motifs typical of densoviruses. Triangles indicate positions of amino acid motifs. Consensus sequences above sequence logos are defined by a 90% identity agreement among all sea star-associated densoviruses.
FIG 5. An overview of endogenized densovirus elements (EVEs) illustrating the conserved existence of Walker box ATPase motifs. EVEs found in this study are shown in red. Sequences in alignment are grouped by host phylum. Sequences are labeled by host species and by origin of sequence from host genome (G), extant virus (V), and host transcriptome (T) and NCBI accession number. Amino acids in bold indicate a sequence identity of 75% or greater within the alignment. The blue highlighted region indicates the NTP-binding and helicase region containing Walker A, B, and C motifs found within the NS1/Rep ORF in densoviruses and parvoviruses.
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