title: Trypanosoma cruzi: gene expression surveyed by proteomic analysis reveals interaction between different genotypes in mixed in vitro cultures.
abstract: We have analyzed the comportment in in vitro culture of 2 different genotypes of Trypanosoma cruzi, the agent of Chagas disease, pertaining to 2 major genetic subdivisions (near-clades) of this parasite. One of the stocks was a fast-growing one, highly virulent in mice, while the other one was slow-growing, mildly virulent in mice. The working hypothesis was that mixtures of genotypes interact, a pattern that has been observed by us in empirical experimental studies. Genotype mixtures were followed every 7 days and characterized by the DIGE technology of proteomic analysis. Proteic spots of interest were characterized by the SAMESPOT software. Patterns were compared to those of pure genotypes that were also evaluated every 7 days. One hundred and three spots exhibited changes in time by comparison with T = 0. The major part of these spots (58%) exhibited an under-expression pattern by comparison with the pure genotypes. 32% of the spots were over-expressed; 10% of spots were not different from those of pure genotypes. Interestingly, interaction started a few minutes after the mixtures were performed. We have retained 43 different proteins that clearly exhibited either under- or over-expression. Proteins showing interaction were characterized by mass spectrometry (MALDI-TOF). Close to 50% of them were either tubulins or heat shock proteins. This study confirms that mixed genotypes of T. cruzi interact at the molecular level. This is of great interest because mixtures of genotypes are very frequent in Chagas natural cycles, both in insect vectors and in mammalian hosts, and may play an important role in the transmission and severity of Chagas disease. The methodology proposed here is potentially applicable to any micropathogen, including fungi, bacteria and viruses. It should be of great interest in the case of bacteria, for which the epidemiological and clinical consequences of mixed infections could be underestimated.
authors.affiliation: Unité Mixte de Recherche, Institut de Recherche pour le Développement/Centre National de la Recherche Scientifique/Universités Montpellier 1 and 2, Génétique et Evolution des Maladies Infectieuses, n° 5290, Maladies Infectieuses et Vecteurs Ecologie, Génétique, Evolution et Contrôle (MIVEGEC) Institut de Recherche pour le Développement, Montpellier, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Machin
authors.affiliation: Unité Mixte de Recherche, Institut de Recherche pour le Développement/Centre National de la Recherche Scientifique/Universités Montpellier 1 and 2, Génétique et Evolution des Maladies Infectieuses, n° 5290, Maladies Infectieuses et Vecteurs Ecologie, Génétique, Evolution et Contrôle (MIVEGEC) Institut de Recherche pour le Développement, Montpellier, France.
authors.forename: Jenny
authors.initials: J
authors.lastname: Telleria
authors.affiliation: Unité Mixte de Recherche, Institut de Recherche pour le Développement/Centre National de la Recherche Scientifique, n° 5096, Centre Institut de Recherche pour le Développement, Montpellier, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Brizard
authors.affiliation: Plate-forme de Proteomique Fonctionnelle, c/o Institut de Génomique Fonctionnelle, Centre National de la Recherche Scientifique Unité Mixte de Recherche 5203, Institut National de la Santé et de la Recherche Médicales, Unité 661, Université Montpellier I et II, Montpellier, France.
authors.forename: Edith
authors.initials: E
authors.lastname: Demettre
authors.affiliation: Plate-forme de Proteomique Fonctionnelle, c/o Institut de Génomique Fonctionnelle, Centre National de la Recherche Scientifique Unité Mixte de Recherche 5203, Institut National de la Santé et de la Recherche Médicales, Unité 661, Université Montpellier I et II, Montpellier, France.
authors.forename: Martial
authors.initials: M
authors.lastname: Séveno
authors.affiliation: Department of Ecology and Evolutionary Biology, University of California Irvine, Irvine, California, United States of America.
authors.forename: Francisco José
authors.initials: FJ
authors.lastname: Ayala
authors.affiliation: Unité Mixte de Recherche, Institut de Recherche pour le Développement/Centre National de la Recherche Scientifique/Universités Montpellier 1 and 2, Génétique et Evolution des Maladies Infectieuses, n° 5290, Maladies Infectieuses et Vecteurs Ecologie, Génétique, Evolution et Contrôle (MIVEGEC) Institut de Recherche pour le Développement, Montpellier, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Tibayrenc
mesh_terms: D005838:Genotype; D020543:Proteome; D040901:Proteomics / Q000379:methods; D059467:Transcriptome; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000378:metabolism*
chemical_list: D020543:Proteome
references.citation: C R Acad Sci III. 1985;300(15):555-7
references.pmid: 3926253
references.citation: Proc Natl Acad Sci U S A. 1990 Apr;87(7):2414-8
references.pmid: 2320563
references.citation: Proc Natl Acad Sci U S A. 2010 Nov 23;107(47):20411-6
references.pmid: 21059959
references.citation: PLoS Pathog. 2007 Nov;3(11):e176
references.pmid: 18020704
references.citation: Exp Parasitol. 2006 Apr;112(4):237-46
references.pmid: 16406355
references.citation: Trans R Soc Trop Med Hyg. 1997 Sep-Oct;91(5):602-5
references.pmid: 9463681
references.citation: Parasitology. 1997 Mar;114 ( Pt 3):213-8
references.pmid: 9075341
references.citation: J Cell Sci. 2001 Aug;114(Pt 15):2723-33
references.pmid: 11683407
references.citation: Anal Chem. 1996 Mar 1;68(5):850-8
references.pmid: 8779443
references.citation: Proc Natl Acad Sci U S A. 1986 Jan;83(1):115-9
references.pmid: 3510428
references.citation: Parasitol Res. 1998 May;84(5):348-53
references.pmid: 9610630
references.citation: Infect Genet Evol. 2012 Mar;12(2):240-53
references.pmid: 22226704
references.citation: Annu Rev Genet. 1999;33:449-77
references.pmid: 10690415
references.citation: Exp Parasitol. 1998 May;89(1):30-9
references.pmid: 9603486
references.citation: Trends Parasitol. 2013 Jun;29(6):264-9
references.pmid: 23602631
references.citation: Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):E3305-13
references.pmid: 22949662
references.citation: Anal Chem. 1988 Oct 15;60(20):2299-301
references.pmid: 3239801
references.citation: Science. 2001 May 11;292(5519):1099-102
references.pmid: 11352063