Differential Protein Expressions in Virus-Infected and Uninfected Trichomonas vaginalis
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Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected (V+) and uninfected (V-) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V+ compared with V- isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V+ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V+ and V- isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.
Abstract
Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected (V+) and uninfected (V-) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V+ compared with V- isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V+ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V+ and V- isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.
참고문헌 (32)
- Johnston VJ, "Global epidemiology and control of Trichomonas vaginalis", Curr Opin Infect Dis, 21, pp.56~64 (2008)
- Mielczarek E, "Trichomonas vaginalis : pathogenicity and potential role in human reproductive failure", Infection, 44, pp.447~458 (2016)
- Centers for Disease Control and Prevention, "Sexually transmitted diseases treatment guidelines, 2006", MMWR Recomm Rep, 55, pp.1~94 (2006)
- McClelland RS, "Infection with Trichomonas vaginalis increases the risk of HIV-1 acquisition", J Infect Dis, 195, pp.698~702 (2007)
- ViikkiM, "Gynaecological infections as risk determinants of subsequent cervical neoplasia", Acta Oncol, 39, pp.71~75 (2000)
- Embley TM, "Eukaryotic evolution, changes and challenges", Nature, 440, pp.623~630 (2006)
- Wang A, "Trichomonas vaginalis phenotypic variation occurs only among trichomonads infected with the double-stranded RNA virus", J Exp Med, 166, pp.142~150 (1987)
- Wang AL, "The double-stranded RNA in Trichomonas vaginalis may originate from virus-like particles", Proc Natl Acad Sci U S A, 83, pp.7956~7960 (1986)
- Miller RL, "Purification and characterization of the Giardia lamblia double-stranded RNA virus", Mol Biochem Parasitol, 28, pp.189~195 (1988)
- Stuart KD, "Molecular organization of Leishmania RNA virus 1", Proc Natl Acad Sci U S A, 89, pp.8596~8600 (1992)
- Kniel KE, "Characterization and potential use of a Cryptosporidium parvum virus (CPV) antigen for detecting C. parvum oocysts", J Microbiol Methods, 58, pp.189~195 (2004)
- Khoshnan A, "Trichomonas vaginalis with a double-stranded RNA virus has upregulated levels of phenotypically variable immunogen mRNA", J Virol, 68, pp.4035~4038 (1994)
- Choumet V, "The salivary glands and saliva of Anopheles gambiae as an essential step in the Plasmodium life cycle : a global proteomic study", Proteomics, 7, pp.3384~3394 (2007)
- Lingdan L, "Differential dissolved protein expression throughout the life cycle of Giardia lamblia", Exp Parasitol, 132, pp.465~469 (2012)
- Li W, "The viral RNA-based transfection of enhanced green fluorescent protein(EGFP)in the parasitic protozoan Trichomonas vaginalis", Parasitol Res, 110, pp.1305~1310 (2012)
- Diamond LS, "The establishment of various trichomonads of animals and man in axenic cultures", J Parasitol, 43, pp.488~490 (1957)
- Alderete JF, "Trichomonas vaginalis NYH286 phenotypic variation may be coordinated for a repertoire of trichomonad surface immunogens", Infect Immun, 55, pp.1957~1962 (1987)
- Alderete JF, "Phenotype and protein-epitope phenotypic variation among fresh isolates of Trichomonas vaginalis", Infect Immun, 55, pp.1037~1041 (1987)
- Alderete JF, "Phenotypic variation and diversity among Trichomonas vaginalis isolates and correlation of phenotype with trichomonal virulence determinants", Infect Immun, 53, pp.285~293 (1986)
- Leitsch D, "Down-regulation of flavin reductase and alcohol dehydrogenase-1(ADH1)in metronidazole-resistant isolates of Trichomonas vaginalis", Mol Biochem Parasitol, 183, pp.177~183 (2012)
- Leitsch D, "The flavin inhibitor diphenyleneiodonium renders Trichomonas vaginalis resistant to metronidazole, inhibits thioredoxin reductase and flavin reductase, and shuts off hydrogenosomal enzymatic pathways", Mol Biochem Parasitol, 171, pp.17~24 (2010)
- Snipes LJ, "Molecular epidemiology of metronidazole resistance in a population of Trichomonas vaginalis clinical isolates", J Clin Microbiol, 38, pp.3004~3009 (2000)
- Chen ZQ, "The essential vertebrate ABCE1 protein interacts with eukaryotic initiation factors", J Biol Chem, 281, pp.7452~7457 (2006)
- Mäser P, "Identification of three ABC transporter genes in Trypanosoma brucei spp", Parasitol Res, 84, pp.106~111 (1998)
- Engbring JA, "Three genes encode distinct AP33 proteins involved in Trichomonas vaginalis cytoadherence", Mol Microbiol, 28, pp.305~313 (1998)
- Engbringtand JA, "Characterizationof Trichomonas vaginalis AP33 adhesin and cell surface interactive domains", Microbiology, 144, pp.3011~3018 (1998)
- Garcia AF, "Characterization of the Trichomonas vaginalis surface-associated AP65 and binding domain interacting with trichomonads and host cells", BMC Microbiol, 7, pp.116~ (2007)
- Cimarelli A, "Translation elongation factor 1-alpha interacts specifically with the human immunodeficiency virus type 1 Gag polyprotein", J Virol, 73, pp.5388~5401 (1999)
- Das T, "RNA polymerase of vesicular stomatitis virus specifically associates with translation elongation factor-1αβγ for its activity", Proc Natl Acad Sci USA, 95, pp.1449~1454 (1998)
- Fujiki R, "GlcNAcylation of histone H2B facilitates its monoubiquitination", Nature, 480, pp.557~560 (2011)
- Hartl FU, "Molecular chaperones in the cytosol : from nascent chain to folded protein", Science, 295, pp.1852~1858 (2002)
- Aparicio F, "Virus induction of heat shock protein 70 reflects a general response to protein accumulation in the plant cytosol", Plant Physiol, 138, pp.529~536 (2005)