Dataset Information

A multiplex targeted mass spectrometry assay for one-shot flavivirus diagnosis

ABSTRACT: Targeted proteomic mass spectrometry is emerging as a salient clinical diagnostic tool to track protein biomarkers. Yet, its strong analytical properties have not been exploited in the diagnosis and typing of flavivirus. Here, we report the development of a highly sensitive and specific single-shot robust assay for flavivirus typing and diagnosis using targeted mass spectrometry technology. Our flavivirus parallel reaction monitoring assay (fvPRM) has the ability to track secreted flaviviral non-structural protein 1 (NS1) over a broad diagnostic and typing window with high sensitivity, specificity, extendibility and multiplexing capability. These features, pivotal and pertinent to efficient response towards flavivirus outbreaks, including newly emerging flavivirus strains, and circumvent the limitations of current diagnostic assays. fvPRM thus carries high potential in positioning itself as a forerunner in delivering early and accurate diagnosis for disease management.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Flavivirus

TISSUE(S): Hepatocyte, Blood Plasma, Blood Serum

SUBMITTER: Sheena Wee

LAB HEAD: Jayantha Gunaratne

PROVIDER: PXD006922 | Pride | 2019-03-25

REPOSITORIES: Pride

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Publications

Multiplex targeted mass spectrometry assay for one-shot flavivirus diagnosis.

Wee Sheena S Alli-Shaik Asfa A Kek Relus R Swa Hannah L F HLF Tien Wei-Ping WP Lim Vanessa W VW Leo Yee-Sin YS Ng Lee-Ching LC Hapuarachchi Hapuarachchige C HC Gunaratne Jayantha J

Proceedings of the National Academy of Sciences of the United States of America 20190318 14

Targeted proteomic mass spectrometry is emerging as a salient clinical diagnostic tool to track protein biomarkers. However, its strong analytical properties have not been exploited in the diagnosis and typing of flaviviruses. Here, we report the development of a sensitive and specific single-shot robust assay for flavivirus typing and diagnosis using targeted mass spectrometry technology. Our flavivirus parallel reaction monitoring assay (<i>fv</i>PRM) has the ability to track secreted flavivir ...[more]

PMID: 30886083

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Project description:The high osmolarity glycerol response (HOG) pathway plays a pivotal role in the stress response, virulence regulation, and differentiation of fungi, including Cryptococcus neoformans that causes fatal meningoencephalitis. Core signaling components of in the HOG pathway, including the Tco-Ypd1-Ssk1 phosphorelay system and the Ssk2-Pbs2-Hog1 MAPK module, have been elucidated but its downstream transcription factors remain unclear. Here we demonstrated that Atf1 with a basic leucine zipper domain is the transcription factor downstream of Hog1 in C. neoformans. We found that ATF1 expression was differentially regulated by oxidative damaging agents, mainly in a Hog1-dependent, but Mpk1-independent, manner. Interestingly, Atf1 not only promoted oxidative stress response and adaptation, but also played an opposing role to Hog1 in the process. Atf1 primarily localized to the nucleus under both unstressed and oxidative stress conditions in a Hog1-independent manner. Our data demonstrated that Atf1 promoted pheromone production and sexual differentiation under negative control by Hog1. Finally, a DNA microarray-based transcriptome analysis of the atf1M-bM-^HM-^F mutant under unstressed and oxidative stress conditions revealed that Atf1 regulated oxidative stress response genes, including a sulfiredoxin gene (SRX1). Intriguing, the array data further demonstrated that Atf1 modulated basal expression of genes involved in DNA repair and genotoxic stress response. Supporting this, we found that the atf1M-bM-^HM-^F mutant was highly sensitive to genotoxic agents. In conclusion, this study provided a further insight into the Hog1-dependent oxidative and genotoxic stress response and differentiation mechanism of C. neoformans. There are more than 95% of genome homology between JEC21 and H99. Therefore 6 slides of JEC21 (Cryptococcus neoformans var. neoformans serotype D) 70-mer oligo are used in this analysis, 3 biological replicate experiments are performed, total RNAs are extracted 2 conditions (with or without treatment of Hydroten peroxide) from H99 (H99 Wild type strain (Cryptococcus neoformans var. grubii serotype A), and atf1M-NM-^T). We use the mixed all of total RNAs from this experiment as a control RNA. We use Cy3 as Sample dye and Cy5 as a control dye.

Dataset Information

A multiplex targeted mass spectrometry assay for one-shot flavivirus diagnosis

Publications

Multiplex targeted mass spectrometry assay for one-shot flavivirus diagnosis.

Similar Datasets

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