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Trypanin Disruption Affects the Motility and Infectivity of the Protozoan Trypanosoma cruzi.
Saenz-Garcia, Jose L; Borges, Beatriz S; Souza-Melo, Normanda; Machado, Luiz V; Miranda, Juliana S; Pacheco-Lugo, Lisandro Alfonso; Moretti, Nilmar S; Wheleer, Richard; Soares Medeiros, Lia C; DaRocha, Wanderson D.
Afiliação
  • Saenz-Garcia JL; Laboratório de Genômica Funcional de Parasitos (GFP), Universidade Federal de Paraná, Curitiba, Brazil.
  • Borges BS; Laboratório de Biologia Celular, Instituto Carlos Chagas, Fundação Oswaldo Cruz (Fiocruz), Curitiba, Brazil.
  • Souza-Melo N; Laboratório de Biologia Molecular de Patógenos (LBMP), Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
  • Machado LV; Laboratório de Ultraestrutura Hertha Mayer, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
  • Miranda JS; Laboratório de Genômica Funcional de Parasitos (GFP), Universidade Federal de Paraná, Curitiba, Brazil.
  • Pacheco-Lugo LA; Laboratório de Genômica Funcional de Parasitos (GFP), Universidade Federal de Paraná, Curitiba, Brazil.
  • Moretti NS; Facultad de Ciencias Básicas y Biomédicas, Universidad Simón Bolívar, Barranquilla, Colombia.
  • Wheleer R; Laboratório de Biologia Molecular de Patógenos (LBMP), Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
  • Soares Medeiros LC; Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
  • DaRocha WD; Laboratório de Biologia Celular, Instituto Carlos Chagas, Fundação Oswaldo Cruz (Fiocruz), Curitiba, Brazil.
Front Cell Infect Microbiol ; 11: 807236, 2021.
Article em En | MEDLINE | ID: mdl-35071054
The flagellum of Trypanosomatids is an organelle that contributes to multiple functions, including motility, cell division, and host-pathogen interaction. Trypanin was first described in Trypanosoma brucei and is part of the dynein regulatory complex. TbTrypanin knockdown parasites showed motility defects in procyclic forms; however, silencing in bloodstream forms was lethal. Since TbTrypanin mutants show drastic phenotypic changes in mammalian stages, we decided to evaluate if the Trypanosoma cruzi ortholog plays a similar role by using the CRISPR-Cas9 system to generate null mutants. A ribonucleoprotein complex of SaCas9 and sgRNA plus donor oligonucleotide were used to edit both alleles of TcTrypanin without any selectable marker. TcTrypanin -/- epimastigotes showed a lower growth rate, partially detached flagella, normal numbers of nuclei and kinetoplasts, and motility defects such as reduced displacement and speed and increased tumbling propensity. The epimastigote mutant also showed decreased efficiency of in-vitro metacyclogenesis. Mutant parasites were able to complete the entire life cycle in vitro; however, they showed a reduction in their infection capacity compared with WT and addback cultures. Our data show that T. cruzi life cycle stages have differing sensitivities to TcTrypanin deletion. In conclusion, additional work is needed to dissect the motility components of T. cruzi and to identify essential molecules for mammalian stages.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Trypanosoma brucei brucei / Trypanosoma cruzi / Doença de Chagas Limite: Animals Idioma: En Revista: Front Cell Infect Microbiol Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Brasil País de publicação: Suíça
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Trypanosoma brucei brucei / Trypanosoma cruzi / Doença de Chagas Limite: Animals Idioma: En Revista: Front Cell Infect Microbiol Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Brasil País de publicação: Suíça