title: In-depth proteomic analysis of Varroa destructor: Detection of DWV-complex, ABPV, VdMLV and honeybee proteins in the mite.
abstract: We investigated pathogens in the parasitic honeybee mite Varroa destructor using nanoLC-MS/MS (TripleTOF) and 2D-E-MS/MS proteomics approaches supplemented with affinity-chromatography to concentrate trace target proteins. Peptides were detected from the currently uncharacterized Varroa destructor Macula-like virus (VdMLV), the deformed wing virus (DWV)-complex and the acute bee paralysis virus (ABPV). Peptide alignments revealed detection of complete structural DWV-complex block VP2-VP1-VP3, VDV-1 helicase and single-amino-acid substitution A/K/Q in VP1, the ABPV structural block VP1-VP4-VP2-VP3 including uncleaved VP4/VP2, and VdMLV coat protein. Isoforms of viral structural proteins of highest abundance were localized via 2D-E. The presence of all types of capsid/coat proteins of a particular virus suggested the presence of virions in Varroa. Also, matches between the MWs of viral structural proteins on 2D-E and their theoretical MWs indicated that viruses were not digested. The absence/scarce detection of non-structural proteins compared with high-abundance structural proteins suggest that the viruses did not replicate in the mite; hence, virions accumulate in the Varroa gut via hemolymph feeding. Hemolymph feeding also resulted in the detection of a variety of honeybee proteins. The advantages of MS-based proteomics for pathogen detection, false-positive pathogen detection, virus replication, posttranslational modifications, and the presence of honeybee proteins in Varroa are discussed.
authors.affiliation: Crop Research Institute, Prague 6, Czechia.
authors.forename: Tomas
authors.initials: T
authors.lastname: Erban
authors.affiliation: Laboratory of Mass Spectrometry, Charles University in Prague, Faculty of Science, Prague 2, Czechia.
authors.forename: Karel
authors.initials: K
authors.lastname: Harant
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Prague 6, Czechia.
authors.forename: Martin
authors.initials: M
authors.lastname: Hubalek
authors.affiliation: Crop Research Institute, Prague 6, Czechia.
authors.forename: Pavel
authors.initials: P
authors.lastname: Vitamvas
authors.affiliation: Bee Research Institute at Dol, Libcice nad Vltavou, Czechia.
authors.forename: Martin
authors.initials: M
authors.lastname: Kamler
authors.affiliation: CNR ISPA, Agrofood Dept, I-73100 Lecce, Italy.
authors.forename: Palmiro
authors.initials: P
authors.lastname: Poltronieri
authors.affiliation: Bee Research Institute at Dol, Libcice nad Vltavou, Czechia.
authors.forename: Jan
authors.initials: J
authors.lastname: Tyl
authors.affiliation: Crop Research Institute, Prague 6, Czechia.
authors.forename: Martin
authors.initials: M
authors.lastname: Markovic
authors.affiliation: Bee Research Institute at Dol, Libcice nad Vltavou, Czechia.
authors.forename: Dalibor
authors.initials: D
authors.lastname: Titera
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D030541:Databases, Genetic; D054884:Host-Pathogen Interactions; D020543:Proteome; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry; D057077:Varroidae / Q000821:virology*
chemical_list: D020543:Proteome