Heterologous Expression of the Pathogen-Specific LIC11711 Gene in the Saprophyte L. biflexa Increases Bacterial Binding to Laminin and Plasminogen

Autor(es)

Kochi, Leandro Toshio ^{[1, 2]} ; Fernandes, Luis Guilherme Virgilio ^[2] ; Nascimento, Ana Lucia Tabet Oller ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Ciencias Biomed, BR-05508900 Sao Paulo - Brazil

^[2] Inst Butantan, Lab Desenvolvimento Vacinas, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PATHOGENS; v. 9, n. 8 AUG 2020.
• Citações Web of Science: 0

Resumo

Leptospirosis is a febrile disease and the etiological agents are pathogenic bacteria of the genus Leptospira. The leptospiral virulence mechanisms are not fully understood and the application of genetic tools is still limited, despite advances in molecular biology techniques. The leptospiral recombinant protein LIC11711 has shown interaction with several host components, indicating a potential function in virulence. This study describes a system for heterologous expression of the L. interrogans gene lic11711 using the saprophyte L. biflexa serovar Patoc as a surrogate, aiming to investigate its possible activity in bacterial virulence. Heterologous expression of LIC11711 was performed using the pMaOri vector under regulation of the lipL32 promoter. The protein was found mainly on the leptospiral outer surface, confirming its location. The lipL32 promoter enhanced the expression of LIC11711 in L. biflexa compared to the pathogenic strain, indicating that this strategy may be used to overexpress low-copy proteins. The presence of LIC11711 enhanced the capacity of L. biflexa to adhere to laminin (Lam) and plasminogen (Plg)/plasmin (Pla) in vitro, suggesting the involvement of this protein in bacterial pathogenesis. We show for the first time that the expression of LIC11711 protein of L. interrogans confers a virulence-associated phenotype on L. biflexa, pointing out possible mechanisms used by pathogenic leptospires. (AU)

• Processo FAPESP: 18/09652-4 - Avaliação da interação de proteínas de Leptospira interrogans aos componentes do hospedeiro por meio da construção de Leptospira biflexa recombinante
• Beneficiário: Leandro Toshio Kochi
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 17/06731-8 - Aplicação de CRISPR-interferência na elucidação da patogênese da leptospirose e desenvolvimento de novas estratégias para obtenção de mutantes knockout
• Beneficiário: Luis Guilherme Virgílio Fernandes
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/50981-0 - Busca de proteínas de superfície nas sequências do genoma da Leptospira interrogans: caracterização funcional e imunológica para o entendimento de mecanismos envolvidos na patogênese de bactéria
• Beneficiário: Ana Lucia Tabet Oller do Nascimento
• Modalidade de apoio: Auxílio à Pesquisa - Temático