title: A proteomics approach reveals molecular manipulators of distinct cellular processes in the salivary glands of Glossina m. morsitans in response to Trypanosoma b. brucei infections.
abstract: Glossina m. morsitans is the primary vector of the Trypanosoma brucei group, one of the causative agents of African trypanosomoses. The parasites undergo metacyclogenesis, i.e. transformation into the mammalian-infective metacyclic trypomastigote (MT) parasites, in the salivary glands (SGs) of the tsetse vector. Since the MT-parasites are largely uncultivable in vitro, information on the molecular processes that facilitate metacyclogenesis is scanty.
  
  
  
  To bridge this knowledge gap, we employed tandem mass spectrometry to investigate protein expression modulations in parasitized (T. b. brucei-infected) and unparasitized SGs of G. m. morsitans. We annotated the identified proteins into gene ontologies and mapped the up- and downregulated proteins within protein-protein interaction (PPI) networks.
  
  
  
  We identified 361 host proteins, of which 76.6 % (n = 276) and 22.3 % (n = 81) were up- and downregulated, respectively, in parasitized SGs compared to unparasitized SGs. Whilst 32 proteins were significantly upregulated (> 10-fold), only salivary secreted adenosine was significantly downregulated. Amongst the significantly upregulated proteins, there were proteins associated with blood feeding, immunity, cellular proliferation, homeostasis, cytoskeletal traffic and regulation of protein turnover. The significantly upregulated proteins formed major hubs in the PPI network including key regulators of the Ras/MAPK and Ca(2+)/cAMP signaling pathways, ubiquitin-proteasome system and mitochondrial respiratory chain. Moreover, we identified 158 trypanosome-specific proteins, notable of which were proteins in the families of the GPI-anchored surface glycoproteins, kinetoplastid calpains, peroxiredoxins, retrotransposon host spot multigene and molecular chaperones. Whilst immune-related trypanosome proteins were over-represented, membrane transporters and proteins involved in translation repression (e.g. ribosomal proteins) were under-represented, potentially reminiscent of the growth-arrested MT-parasites.
  
  
  
  Our data implicate the significantly upregulated proteins as manipulators of diverse cellular processes in response to T. b. brucei infection, potentially to prepare the MT-parasites for invasion and evasion of the mammalian host immune defences. We discuss potential strategies to exploit our findings in enhancement of trypanosome refractoriness or reduce the vector competence of the tsetse vector.
authors.affiliation: Biotechnology Research Institute, Kenya Agricultural and Livestock Research Organization, P.O Box 57811, 00200, Kaptagat Rd, Loresho, Nairobi, Kenya. henry.kariithi@kalro.org.|Insect Pest Control Laboratories, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency, Wagrammer Straße 5, Vienna, Austria. henry.kariithi@kalro.org.
authors.forename: Henry M
authors.initials: HM
authors.lastname: Kariithi
authors.affiliation: Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703, HA, Wageningen, The Netherlands.
authors.forename: Sjef
authors.initials: S
authors.lastname: Boeren
authors.affiliation: Department of Biochemistry and Molecular Biology, Egerton University, P.O. Box 536, 20115, Njoro, Kenya.
authors.forename: Edwin K
authors.initials: EK
authors.lastname: Murungi
authors.affiliation: Laboratory of Virology, Wageningen University, Droevendaalsesteeg 1, 6708, PB, Wageningen, The Netherlands.
authors.forename: Just M
authors.initials: JM
authors.lastname: Vlak
authors.affiliation: Insect Pest Control Laboratories, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, International Atomic Energy Agency, Wagrammer Straße 5, Vienna, Austria. a.m.m.abd-alla@iaea.org.
authors.forename: Adly M M
authors.initials: AM
authors.lastname: Abd-Alla
mesh_terms: D000818:Animals; D006790:Host-Parasite Interactions; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D040901:Proteomics; D012469:Salivary Glands / Q000737:chemistry / Q000469:parasitology; D014346:Trypanosoma brucei brucei / Q000254:growth & development / Q000502:physiology*; D014370:Tsetse Flies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000469:parasitology*
chemical_list: D019476:Insect Proteins
keywords: LC-MS/MS; Metacyclic trypomastigotes; Metacyclogenesis; Protein-protein interaction; Trypanosome refractoriness; Vector competence