iVirus 2.0: Cyberinfrastructure-supported tools and data to power DNA virus ecology

Microbes drive myriad ecosystem processes, but under strong influence from viruses. Because studying viruses in complex systems requires different tools than those for microbes, they remain underexplored. To combat this, we previously aggregated double-stranded DNA (dsDNA) virus analysis capabilities and resources into ‘iVirus’ on the CyVerse collaborative cyberinfrastructure. Here we substantially expand iVirus’s functionality and accessibility, to iVirus 2.0, as follows. First, core iVirus apps were integrated into the Department of Energy’s Systems Biology KnowledgeBase (KBase) to provide an additional analytical platform. Second, at CyVerse, 20 software tools (apps) were upgraded or added as new tools and capabilities. Third, nearly 20-fold more sequence reads were aggregated to capture new data and environments. Finally, documentation, as “live” protocols, was updated to maximize user interaction with and contribution to infrastructure development. Together, iVirus 2.0 serves as a uniquely central and accessible analytical platform for studying how viruses, particularly dsDNA viruses, impact diverse microbial ecosystems.

Draft Genome Sequence of Sicyoidochytrium minutum DNA Virus Strain 001

Sicyoidochytrium minutum DNA virus strain 001 (SmDNAV 001) is a double-stranded DNA (dsDNA) virus that infects the marine fungoid protist Sicyoidochytrium minutum . We report the draft genome sequence of SmDNAV 001. The 236,345-bp genome contained 358 coding sequences (CDSs) and three tRNA-coding sequences.

Tracing dsDNA Virus–Host Coevolution through Correlation of Their G-Quadruplex-Forming Sequences

The importance of gene expression regulation in viruses based upon G-quadruplex may point to its potential utilization in therapeutic targeting. Here, we present analyses as to the occurrence of putative G-quadruplex-forming sequences (PQS) in all reference viral dsDNA genomes and evaluate their dependence on PQS occurrence in host organisms using the G4Hunter tool. PQS frequencies differ across host taxa without regard to GC content. The overlay of PQS with annotated regions reveals the localization of PQS in specific regions. While abundance in some, such as repeat regions, is shared by all groups, others are unique. There is abundance within introns of Eukaryota-infecting viruses, but depletion of PQS in introns of bacteria-infecting viruses. We reveal a significant positive correlation between PQS frequencies in dsDNA viruses and corresponding hosts from archaea, bacteria, and eukaryotes. A strong relationship between PQS in a virus and its host indicates their close coevolution and evolutionarily reciprocal mimicking of genome organization.

Virus-host coexistence in phytoplankton through the genomic lens

Virus-microbe interactions in the ocean are commonly described by “boom and bust” dynamics, whereby a numerically dominant microorganism is lysed and replaced by a virus-resistant one. Here, we isolated a microalga strain and its infective dsDNA virus whose dynamics are characterized instead by parallel growth of both the microalga and the virus. Experimental evolution of clonal lines revealed that this viral production originates from the lysis of a minority of virus-susceptible cells, which are regenerated from resistant cells. Whole-genome sequencing demonstrated that this resistant-susceptible switch involved a large deletion on one chromosome. Mathematical modeling explained how the switch maintains stable microalga-virus population dynamics consistent with their observed growth pattern. Comparative genomics confirmed an ancient origin of this “accordion” chromosome despite a lack of sequence conservation. Together, our results show how dynamic genomic rearrangements may account for a previously overlooked coexistence mechanism in microalgae-virus interactions.

Molecular fossils reveal ancient associations of dsDNA viruses with several phyla of fungi

Little is known about the infections of double-stranded DNA (dsDNA) viruses in fungi. Here, we use a paleovirological method to systematically identify the footprints of past dsDNA virus infections within the fungal genomes. We uncover two distinct groups of endogenous nucleocytoplasmic large DNA viruses (NCLDVs) in at least seven fungal phyla (accounting for about a third of known fungal phyla), revealing an unprecedented diversity of dsDNA viruses in fungi. Interestingly, one fungal dsDNA virus lineage infecting six fungal phyla is closely related to the giant virus Pithovirus, suggesting giant virus relatives might widely infect fungi. Co-speciation analyses indicate fungal NCLDVs mainly evolved through cross-species transmission. Taken together, our findings provide novel insights into the diversity and evolution of NCLDVs in fungi.

A57 An evolutionary framework to guide the hunt for human dsDNA viruses

It is becoming increasingly obvious that we only know a fraction of the human virome. The development of next-generation sequencing (NGS) technologies has dramatically increased our ability to hunt viruses. Yet, the small genomes and low copy numbers characteristic of most viruses make undirected (shotgun) hunts a relatively inefficient strategy. Here, we propose to speed-up the rate of double-stranded DNA (dsDNA) virus discovery by combining NGS with evolutionary thinking. dsDNA viruses are thought to have co-diverged with their hosts. As this applies to the hominine lineages (gorillas, humans, chimpanzees, and bonobos), it is theoretically possible to estimate the phylogenetic position of cryptic human viruses by identifying co-divergent viral lineages infecting non-human hominines. Where these lineages do not comprise a human-infecting counterpart, a yet-unknown human virus may be hiding. The first phase of this project will consist in the high-throughput characterization of dsDNA viruses (herpesviruses, papillomaviruses, and polyomaviruses) infecting wild gorillas, chimpanzees, and bonobos. For this, we will use an exhaustive collection of fecal samples (in terms of hominine species/sub-species diversity) and apply a discovery strategy combining in-solution capture and NGS. This strategy has been developed in the ancient DNA field but has a very broad applicability; it will constitute a nice addendum to the institute technical portfolio. Thanks to the massive amount of information collected, we will be able to reconstruct the evolutionary history of many dsDNA virus lineages and to identify those where a human virus would be expected but is still unknown. This will pave the way to the second phase of the project which will consist in a pre-oriented dsDNA human virus hunt based on the use of specific PCR systems implemented in multiplex. We expect that this project will generate an unprecedented amount of data on the processes at play along dsDNA virus evolution (co-divergence versus cross-species transmission), help determine the directionality, frequency, and timing of cross-species transmission events between hominines and unveil the existence of yet-to-be-discovered human viruses.

Insane in the Membrane: Glial Extracellular Vesicles Transmit Polyomaviruses

ABSTRACT Extracellular vesicles (EVs) are major vehicles for transporting viruses en bloc among hosts. While RNA viruses make up the great majority of transmission by EVs, in a recent article in mBio (mBio 10:e00379-19, 2019, https://mbio.asm.org/content/10/2/e00379-19.long), Morris-Love and colleagues revealed that a double-stranded DNA (dsDNA) virus, JC polyomavirus (JCPyV), a major cause of progressive multifocal leukoencephalopathy (PML), can be released from and transmitted to other glia in EVs. This mode of transmission appears to be highly infectious, independent of the free virus attachment and entry receptors LSTc and 5-HT2, and protected from neutralizing antibodies. This novel form of JCPyV transmission may potentially explain its dissemination into the central nervous system (CNS) and its increased virulence.