Gene Gangs of the Chloroviruses: Conserved Clusters of Collinear Monocistronic Genes

Chloroviruses (family Phycodnaviridae) are dsDNA viruses found throughout the world’s inland waters. The open reading frames in the genomes of 41 sequenced chloroviruses (330 + 40 kbp each) representing three virus types were analyzed for evidence of evolutionarily conserved local genomic “contexts”, the organization of biological information into units of a scale larger than a gene. Despite a general loss of synteny between virus types, we informatically detected a highly conserved genomic context defined by groups of three or more genes that we have termed “gene gangs”. Unlike previously described local genomic contexts, the definition of gene gangs requires only that member genes be consistently co-localized and are not constrained by strand, regulatory sites, or intervening sequences (and therefore represent a new type of conserved structural genomic element). An analysis of functional annotations and transcriptomic data suggests that some of the gene gangs may organize genes involved in specific biochemical processes, but that this organization does not involve their coordinated expression

Videokujunduse loomine Vanemuise teatri lavastusele "Rännakud. Maarjamaa laulud"

http://www.ester.ee/record=b5143827*es

CoCAS: a ChIP-on-chip analysis suite

Motivation: High-density tiling microarrays are increasingly used in combination with ChIP assays to study transcriptional regulation. To ease the analysis of the large amounts of data generated by this approach, we have developed ChIP-on-chip Analysis Suite (CoCAS), a standalone software suite which implements optimized ChIP-on-chip data normalization, improved peak detection, as well as quality control reports. Our software allows dye swap, replicate correlation and connects easily with genome browsers and other peak detection algorithms. CoCAS can readily be used on the latest generation of Agilent high-density arrays. Also, the implemented peak detection methods are suitable for other datasets, including ChIP-Seq output

A la redécouverte des Chlorovirus : Contribution à l'étude des virus géants à ADN

Les virus du genre Chlorovirus (phycoDNAviridae) sont des virus à ADN double-brin de grande taille, qui infectent des algues vertes eucaryotes unicellulaires vivant en eau douce nommées les chlorelles (Trebouxiophyceae). A l'aide d'approches bioinformatiques, j'ai consacré mon travail de thèse à l'étude de la diversité génomique des chlorovirus et de leur histoire évolutive, ainsi qu'à l'étude transcriptomique de l'infection virale chez son hôte.Dans le premier volet de ma thèse, j'ai procédé à l'assemblage et à l'annotation de 50 nouveaux génomes de chlorovirus récemment séquencés (Roche-454). J'ai ainsi été en mesure de mieux caractériser les différences de structure du génome et de contenu en gène des différents chlorovirus en fonction de leurs hôtes. J'ai également mis en évidence l'existence d'un quatrième sous-groupe de chlorovirus, repoussant les limites connues de la diversité de ces virus. Enfin j'ai montré que les Chlorovirus ne suivent pas le même schéma évolutif des autres NCLDV et que l'origine de leurs gènes est encore inconnue, bien que probablement virale.Dans un autre projet, j'ai également étudié les variations de la transcription des gènes de la chlorelle (C. varabilis NC64A) induites par l'infection par un chlorovirus (PBCV-1) grâce au séquençage profond (Illumina) des ARNm polyadénylés présent dans la cellule saine, puis après infection. J'ai ainsi pu montrer que les différentes fonctions cellulaires sont impactées de façon préférentielle par l'infection.Giant viruses in the genus Chlorovirus (Phycodnaviridae) infect eukaryotic green microalgae known as Chlorella (Trebouxiophyceae). Using bioinformatic approaches, I dedicated my thesis on the study of the genomic diversity and evolutionnary history of the chlorovirus at the genus level, and the transcriptomic of the viral infection.In the first part on my work, I conducted the assembly and annotation of 50 new chlorovirus genomes recently sequenced (Roche-454). I was able to refine the known differences between chloroviruses, both in genome structure and gene content terms. Clues for the existence of a fourth subgroup of chloroviruses were also found. I was also able to show that the chlorovirus does not follow the same evolutionnary pattern as the other NCLDV, and that the origin of their genes is still largely unknown, but presumably of viral origin.In a second project, I studied the variation in the transcription of chlorella's genes (C. variabilis NC64A) during the infection by a chlorovirus (PBCV-1) using the deep-sequencing (Illumina) of all polyadenylated messenger RNA in the healthy or infected cell. This way, I was able to show that the various cellular functions are preferentially impacted by the infection

Whole-Genome Sequencing of Kaposi’s Sarcoma-Associated Herpesvirus from Zambian Kaposi’s Sarcoma Biopsy Specimens Reveals Unique Viral Diversity

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiological agent for Kaposi’s sarcoma (KS). Both KSHV and KS are endemic in sub-Saharan Africa where approximately 84% of global KS cases occur. Nevertheless, whole-genome sequencing of KSHV has only been completed using isolates from Western countries—where KS is not endemic. The lack of whole-genome KSHV sequence data from the most clinically important geographical region, sub-Saharan Africa, represents an important gap since it remains unclear whether genomic diversity has a role on KSHV pathogenesis. We hypothesized that distinct KSHV genotypes might be present in sub-Saharan Africa compared to Western countries. Using a KSHV-targeted enrichment protocol followed by Illumina deep-sequencing, we generated and analyzed 16 unique Zambian, KS-derived, KSHV genomes. We enriched KSHV DNA over cellular DNA 1,851 to 18,235-fold. Enrichment provided coverage levels up to 24,740-fold; therefore, supporting highly confident polymorphism analysis. Multiple alignment of the 16 newly sequenced KSHV genomes showed low level variability across the entire central conserved region. This variability resulted in distinct phylogenetic clustering between Zambian KSHV genomic sequences and those derived from Western countries. Importantly, the phylogenetic segregation of Zambian from Western sequences occurred irrespective of inclusion of the highly variable genes K1 and K15. We also show that four genes within the more conserved region of the KSHV genome contained polymorphisms that partially, but not fully, contributed to the unique Zambian KSHV whole-genome phylogenetic structure. Taken together, our data suggest that the whole KSHV genome should be taken into consideration for accurate viral characterization

Whole-genome sequencing of KSHV from Zambian Kaposi’s sarcoma biopsies reveals unique viral diversity

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiological agent for Kaposi’s sarcoma (KS). Both KSHV and KS are endemic in sub-Saharan Africa where approximately 84% of global KS cases occur. Nevertheless, whole-genome sequencing of KSHV has only been completed using isolates from Western countries—where KS is not endemic. The lack of whole-genome KSHV sequence data from the most clinically important geographical region, sub-Saharan Africa, represents an important gap as it remains unclear whether genomic diversity has a role on KSHV pathogenesis. We hypothesized that distinct KSHV genotypes might be present in sub-Saharan Africa compared to Western countries. Using a KSHV-targeted enrichment protocol followed by Illumina deep-sequencing, we generated and analyzed sixteen unique Zambian, KS-derived, KSHV genomes. We enriched KSHV DNA over cellular DNA 1,851 to 18,235-fold. Enrichment provided coverage levels up to 24,740-fold; therefore, supporting highly confident polymorphism analysis. Multiple alignment of the sixteen newly sequenced KSHV genomes showed low level variability across the entire central conserved region. This variability resulted in distinct phylogenetic clustering between Zambian KSHV genomic sequences and those derived from Western countries. Importantly, the phylogenetic segregation of Zambian from Western sequences occurred irrespective of inclusion of the highly variable genes K1 and K15. We also show that four genes within the more conserved region of the KSHV genome contained polymorphisms that partially, but not fully, contributed to the unique Zambian KSHV whole-genome phylogenetic structure. Taken together, our data suggest that the whole KSHV genome should be taken into consideration for accurate viral characterization

Gene Gangs of the Chloroviruses: Conserved Clusters of Collinear Monocistronic Genes

Chloroviruses (family Phycodnaviridae) are dsDNA viruses found throughout the world’s inland waters. The open reading frames in the genomes of 41 sequenced chloroviruses (330 + 40 kbp each) representing three virus types were analyzed for evidence of evolutionarily conserved local genomic “contexts”, the organization of biological information into units of a scale larger than a gene. Despite a general loss of synteny between virus types, we informatically detected a highly conserved genomic context defined by groups of three or more genes that we have termed “gene gangs”. Unlike previously described local genomic contexts, the definition of gene gangs requires only that member genes be consistently co-localized and are not constrained by strand, regulatory sites, or intervening sequences (and therefore represent a new type of conserved structural genomic element). An analysis of functional annotations and transcriptomic data suggests that some of the gene gangs may organize genes involved in specific biochemical processes, but that this organization does not involve their coordinated expression

Global Analysis of \u3ci\u3eChlorella variabilis\u3c/i\u3e NC64A mRNA Profiles during the Early Phase of \u3ci\u3eParamecium bursaria\u3c/i\u3e Chlorella Virus-1 Infection

The PBCV-1/Chlorella variabilis NC64A system is a model for studies on interactions between viruses and algae. Here we present the first global analyses of algal host transcripts during the early stages of infection, prior to virus replication. During the course of the experiment stretching over 1 hour, about a third of the host genes displayed significant changes in normalized mRNA abundance that either increased or decreased compared to uninfected levels. The population of genes with significant transcriptional changes gradually increased until stabilizing at 40 minutes post infection. Functional categories including cytoplasmic ribosomal proteins, jasmonic acid biosynthesis and anaphase promoting complex/ cyclosomes had a significant excess in upregulated genes, whereas spliceosomal snRNP complexes and the shikimate pathway had significantly more down-regulated genes, suggesting that these pathways were activated or shut-down in response to the virus infection. Lastly, we examined the expression of C. varibilis RNA polymerase subunits, as PBCV-1 transcription depends on host RNA polymerases. Two subunits were up-regulated, RPB10 and RPC34, suggesting that they may function to support virus transcription. These results highlight genes and pathways, as well as overall trends, for further refinement of our understanding of the changes that take place during the early stages of viral infection

Towards defining the chloroviruses: a genomic journey through a genus of large DNA viruses

Background: Giant viruses in the genus Chlorovirus (family Phycodnaviridae) infect eukaryotic green microalgae. The prototype member of the genus, Paramecium bursaria chlorella virus 1, was sequenced more than 15 years ago, and to date there are only 6 fully sequenced chloroviruses in public databases. Presented here are the draft genome sequences of 35 additional chloroviruses (287 – 348 Kb/319 – 381 predicted protein encoding genes) collected across the globe; they infect one of three different green algal species. These new data allowed us to analyze the genomic landscape of 41 chloroviruses, which revealed some remarkable features about these viruses. Results: Genome colinearity, nucleotide conservation and phylogenetic affinity were limited to chloroviruses infecting the same host, confirming the validity of the three previously known subgenera. Clues for the existence of a fourth new subgenus indicate that the boundaries of chlorovirus diversity are not completely determined. Comparison of the chlorovirus phylogeny with that of the algal hosts indicates that chloroviruses have changed hosts in their evolutionary history. Reconstruction of the ancestral genome suggests that the last common chlorovirus ancestor had a slightly more diverse protein repertoire than modern chloroviruses. However, more than half of the defined chlorovirus gene families have a potential recent origin (after Chlorovirus divergence), among which a portion shows compositional evidence for horizontal gene transfer. Only a few of the putative acquired proteins had close homologs in databases raising the question of the true donor organism(s). Phylogenomic analysis identified only seven proteins whose genes were potentially exchanged between the algal host and the chloroviruses. Conclusion: The present evaluation of the genomic evolution pattern suggests that chloroviruses differ from that described in the related Poxviridae and Mimiviridae. Our study shows that the fixation of algal host genes has been anecdotal in the evolutionary history of chloroviruses. We finally discuss the incongruence between compositional evidence of horizontal gene transfer and lack of close relative sequences in the databases, which suggests that the recently acquired genes originate from a still largely un-sequenced reservoir of genomes, possibly other unknown viruses that infect the same hosts

Global <i>C. variabilis</i> mRNA changes during virus PBCV-1 infection.

<p>(A) Frequency distributions of log₂ abundance ratios for genes between datasets T0 and T7–T60. (B) Frequency of genes with absolute mRNA changes relative to T0>2 fold. (C) Frequency distributions of log₂ abundance ratios for genes between datasets T_n and T_n+20′.</p