Dataset Information

Sequence analysis of Giardia virus proteins by LC-MS/MS

ABSTRACT: Giardiasis, caused by the protozoan parasite Giardia duodenalis, is an intestinal diarrheal disease affecting almost one billion people worldwide. A small endosymbiotic dsRNA viruses, G. lamblia virus (GLV), genus Giardiavirus, family Totiviridae, might inhabit human and animal isolates of G. duodenalis. Three GLV genomes have been sequenced so far, and only one was intensively studied, moreover a positive correlation between GLV and parasite virulence is yet to be proved. To understand the biological significance of GLV infection in Giardia, the characterization of several GLV strains from naturally infected G. duodenalis isolates is necessary. In the contest of high-throughput sequencing of four GLVs strains, from Giardia isolates of human and animal origin, we report on a new, unclassified viral sequence (designed GdRV-2), unrelated to Giardiavirus, encoding and expressing for a single large protein with a RdRp domain homologous to Totiviridae and Botybirnaviridae. We have analyzed the sequence of the GLV capsid protein (CP) and RNA-dependent RNA polymerase (RdRp) by LC-MS/MS analysis using different enzymatic strategies.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Giardia Intestinalis (giardia Lamblia)

SUBMITTER: Serena Camerini

LAB HEAD: Serena Camerini

PROVIDER: PXD025785 | Pride | 2021-07-05

REPOSITORIES: Pride

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Publications

Re-Discovery of Giardiavirus: Genomic and Functional Analysis of Viruses from Giardia duodenalis Isolates.

Marucci Gianluca G Zullino Ilaria I Bertuccini Lucia L Camerini Serena S Cecchetti Serena S Pietrantoni Agostina A Casella Marialuisa M Vatta Paolo P Greenwood Alex D AD Fiorillo Annarita A Lalle Marco M

Biomedicines 20210608 6

Giardiasis, caused by the protozoan parasite Giardia duodenalis, is an intestinal diarrheal disease affecting almost one billion people worldwide. A small endosymbiotic dsRNA viruses, G. lamblia virus (GLV), genus Giardiavirus, family Totiviridae, might inhabit human and animal isolates of G. duodenalis. Three GLV genomes have been sequenced so far, and only one was intensively studied; moreover, a positive correlation between GLV and parasite virulence is yet ...[more]

PMID: 34201207

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Project description:Despite of Giardia duodenalis being one of the most commonly found intestinal pathogens in humans and animals, little is known of the host-parasite interactions in natural hosts. Therefore, the objective of this study was to investigate the intestinal response in calves following a G. duodenalis infection, using a bovine high-density oligo microarray to analyze global gene expression in the small intestine. The resulting microarray data suggested a decrease in inflammation, immune response and immune cell migration in infected animals, which was examined in more detail by quantitative real-time PCR on a panel of cytokines combined with histological analyses. The cytokine transcription levels showed a trend of down regulated expression in infected animals compared to the negative controls, best seen in jejunum for IL-6 and IL-8 and statistically significant for IL-17, IL-13 and IFN-?. No increased immune cell recruitment could be seen after infection, as well as no intestinal pathologies, such as villus shortening or increased levels of apoptosis. Key regulators in this intestinal response seem to be the nuclear peroxisome proliferator-activated receptors alpha (PPARA) and gamma (PPARG), for which an up-regulated expression was seen on microarray and qRT-PCR data. The activation of PPARs can exert an anti-inflammatory effect with inhibition of pro-inflammatory cytokines and a decrease in cell recruitment. . How the PPARs are activated during a Giardia infection still needs to be further elucidated. Eight male Holstein calves aged two to four weeks old were used for the trial. Prior to arrival, all animals were screened for the presence of Giardia cysts in their faecal samples. After confirming their negative status for all these pathogens, four of the animals were randomly chosen and placed in a G. duodenalis contaminated environment, whereas the four remaining animals were kept as negative controls in separate G. duodenalis-free stables. All calves in the study received the same commercial milk replacer. After three weeks, the presence or absence of a G. duodenalis infection was confirmed by IFA on faecal samples after which the animals were euthanized. Changes in gene expression profiles induced by Giardia duodenalis infection were compared using a high-density 60mer bovine oligo microarray.

Dataset Information

Sequence analysis of Giardia virus proteins by LC-MS/MS

Publications

Re-Discovery of <i>Giardiavirus</i>: Genomic and Functional Analysis of Viruses from <i>Giardia duodenalis</i> Isolates.

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