Dataset Information

Proteomics analysis identifies IRSp53 and Fascin as critical for PRV egress and direct cell-cell transmission

ABSTRACT: Pseudorabies virus (PRV) has been widely used as a live transsynaptic tracer for mapping neuronal circuits. Systematically identifying mature PRV virion proteomes and defining copurified host proteins are necessary to fully understand the detailed mechanism underlying PRV transmission processes. Here, we developed a PRV virion purification strategy based on sorting with flow cytometry and characterized the mature extracellular and intracellular PRV virion proteomes using liquid chromatography coupled with tandem mass spectrometry. In addition to viral proteins, a large number of host proteins were also identified, including proteins related to actin cytoskeletal dynamics and membrane protrusion. How many of these host proteins are true virion components are unknown and the majority of these may not be. Through functional analysis, we found that IRSp53 and fascin were critical for the egress process and play a role in direct cell-cell transmission. Moreover, we showed that CDC42 and Rac1 were also involved in the production of mature extracellular virions. Our results suggest that the formation of the filopodia-like cytoskeleton and the rearrangement of the membrane, which are both associated with IRSp53 and fascin, may be important for the transmission of viruses used in neuronal tracing.

ORGANISM(S): Mus Musculus

SUBMITTER: Lin Guo

PROVIDER: PXD015490 | iProX | Wed Sep 18 00:00:00 BST 2019

REPOSITORIES: iProX

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Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell-Cell Transmission.

Yu Fei-Long FL Miao Huan H Xia Jinjin J Jia Fan F Wang Huadong H Xu Fuqiang F Guo Lin L

Proteomics 20191007 23

Pseudorabies virus (PRV) has been widely used as a live trans-synaptic tracer for mapping neuronal circuits. Systematically identifying mature PRV virion proteomes and defining co-purified host proteins are necessary to fully understand the detailed mechanism underlying PRV transmission processes. Here, a PRV virion purification strategy based on sorting with flow cytometry is developed and the mature extracellular and intracellular PRV virion proteomes using LC coupled with MS/MS are characteri ...[more]

PMID: 31531927

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Project description:Plasmodium falciparum gametocyte stages represent a small fraction of the entire parasite biomass that is present during human malaria infection, yet they alone lead to the transmission of this devastating disease. One of the critical gaps in malaria transmission biology and surveillance is our lack of knowledge about gametocyte biology, especially sexual dimorphic development that may influence transmission from the human to the mosquito. Ratios of male and female gametocytes in the peripheral blood can vary significantly; influenced in part by asexual blood stage and gametocyte density as well as vertebrate and invertebrate host factors. Moreover, the role of sex ratios on gametocyte transmission potential to mosquitoes is unknown and dissecting this process has been hampered by the lack of sex-specific protein markers for the circulating, mature stage V gametocytes. The current evidence suggests a high degree of conservation in gametocyte gene complement across Plasmodium, and therefore presumably for sex-specific genes as well. Therefore, to better our understanding of gametocyte development and subsequent infectiousness to mosquitoes, we undertook a two pronged approach. First, we acquired the mixed, male and female stage V gametocyte proteomes of the NF54 isolate and mature stage V female proteome from Dd2, a strain that is defective in producing mature males. Second, we then undertook a Systematic Subtractive Bioinformatic analysis (filtering) approach to identify sex-specific P. falciparum NF54 protein markers based on a comparison with the Dd2 strain and syntenic male and female proteins from the reanalyzed and updated P. berghei (related rodent malaria parasite) gametocyte proteomes. This has produced a short list of putative 174 male- and 258 female-specific P. falciparum stage V proteins. Furthermore, we generated antibodies against three putative female-specific gametocyte stage V proteins in P. falciparum and confirmed sex-specificity for two proteins and also the loss sex-partitioning for a putative female-specific protein in rodent malaria parasites.

Dataset Information

Proteomics analysis identifies IRSp53 and Fascin as critical for PRV egress and direct cell-cell transmission

Publications

Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell-Cell Transmission.

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