title: Polyamine depletion down-regulates expression of the Trichomonas vaginalis cytotoxic CP65, a 65-kDa cysteine proteinase involved in cellular damage.
abstract: Recently, we found that inhibition of putrescine synthesis by ornithine decarboxylase (ODC) significantly increased Trichomonas vaginalis adherence mediated by protein adhesins. Surprisingly and unexpectedly, trichomonal contact-dependent cytotoxicity was absent. Therefore, a role for polyamine depletion on regulation of T. vaginalis cytotoxicity mediated by the cysteine proteinase (CP) of 65-kDa, CP65, was investigated. We performed cytotoxicity and cell-binding assays followed by zymograms, as well as Western blot and indirect immunofluorescence assays using specific anti-CP65 antibodies to detect CP65. Trichomonads grown in the presence of the ODC inhibitor, 1-4-diamino-2-butanone (DAB) had lower levels of cytotoxicity that corresponded with diminished CP65 proteolytic activity when compared to untreated organisms handled identically. Likewise, semiquantitative and qRT-PCR as well as Western blot and immunofluorescence assays showed decreased amounts of tvcp65 mRNA and CP65 protein in DAB-treated parasites. These effects were reversed by addition of exogenous putrescine. These data show a direct link between polyamine metabolism and expression of the cytotoxic CP65 proteinase involved in trichomonal host cellular damage.
authors.affiliation: Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México (UACM), San Lorenzo # 290, Col. Del Valle, CP 03100 Mexico City, Mexico.
authors.forename: María Elizbeth
authors.initials: ME
authors.lastname: Alvarez-Sánchez
authors.forename: Bertha Isabel
authors.initials: BI
authors.lastname: Carvajal-Gamez
authors.forename: Eduardo
authors.initials: E
authors.lastname: Solano-González
authors.forename: Máximo
authors.initials: M
authors.lastname: Martínez-Benitez
authors.forename: Ana F
authors.initials: AF
authors.lastname: Garcia
authors.forename: John F
authors.initials: JF
authors.lastname: Alderete
authors.forename: Rossana
authors.initials: R
authors.lastname: Arroyo
mesh_terms: D000818:Animals; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*; D015536:Down-Regulation; D006801:Humans; D011073:Polyamines / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*; D014245:Trichomonas Infections / Q000378:metabolism / Q000473:pathology; D014246:Trichomonas vaginalis / Q000187:drug effects / Q000201:enzymology* / Q000472:pathogenicity
chemical_list: D011073:Polyamines; D015800:Protozoan Proteins; D003546:Cysteine Endopeptidases