Protein structural biology applied in the study of c-di-GMP signaling and bacterial secretion systems

• Grant number: 22/08730-7
• Support Opportunities: Regular Research Grants
• Start date: May 01, 2023
• End date: April 30, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Cristiane Rodrigues Guzzo Carvalho
• Grantee: Cristiane Rodrigues Guzzo Carvalho
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Robson Francisco de Souza

Abstract

In this work, we will be working with two bacteria: the plant pathogen Xanthomonas citri (X. citri) and the human pathogen Leptospira interrogans (Lic). X. citri causes citrus canker in citrus crops, causing significant damage to Brazilian agriculture. It is a bacterium that uses an extracellular appendage, the type IV pilus (T4P), to move around (twitching). This filament is assembled through a protein nanomachine that crosses the bacterial wall, and some bacteriophages, such as ¦Xacm4-11, take advantage of these extracellular structures to infect X. citri. We obtained structural data on the T4P secretin, PilQ, which was co-purified with the TsaP protein. In this regular project, we intend to refine this structure and understand the role of TsaP in T4P function. In addition, our group was able to purify the isolated T4P filament and in complex with ¦Xacm4-11But the complex is not stable, and we need to optimize the purification conditions. Cryo-EM data collection of the filament has been obtained, and we are trying to solve its helical structure. Our research group has shown that X. citri produces outer membrane vesicles (OMVs), and we are investigating their composition and biological role (see manuscript on BioRxiv: https://doi.org/10.1101/2021.04.26.441564).Leptospirosis is an infectious disease caused by species of the bacterial genus Leptospira and is more common in tropical areas with poor sanitation. In Brazil, it is endemic and causes about 1,000 deaths per year. The disease can range from asymptomatic to severe and has a significant impact on the economy, emphasizing the importance of studying the biology and pathogenesis of this neglected pathogen. Our goal is to study the signaling network involving the second messenger c-di-GMP, with a focus on refining the structure of the diguanylate cyclase LIC_11128, which binds to brevianamide F. We do not yet know the effect of this natural compound on the enzymatic activity of LIC_11128 and on the survival of the bacterium. In addition, we will finalize our functional studies of the possible virulence factor LIC_11568, located in the T2SS cluster. Our studies have the potential to provide important information about the biology and pathogenesis of these bacteria, contributing to the understanding and control of diseases that affect agriculture and human health. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

VINCES, TANIA CHURASACARI; DE SOUZA, ANACLETO SILVA; CARVALHO, CECILIA F.; VISNARDI, ALINE BIAZOLA; TEIXEIRA, RAPHAEL D.; LLONTOP, EDGAR E.; BISMARA, BEATRIZ APARECIDA PASSOS; VICENTE, ELISABETE J.; PEREIRA, JOSE O.; DE SOUZA, ROBSON FRANCISCO; et al. Monomeric Esterase: Insights into Cooperative Behavior, Hysteresis/Allokairy. BIOCHEMISTRY, v. 63, n. 9, p. 16-pg., 2024-04-26. (21/10577-0, 19/12234-2, 22/08730-7, 21/05262-0, 19/00195-2, 17/17303-7, 20/04680-0)

VISNARDI, ALINE BIAZOLA; RIBEIRO, RODOLFO ALVARENGA; DE SOUZA, ANACLETO SILVA; VINCES, TANIA GERALDINE CHURASACARI; LLONTOP, EDGAR E.; FERRARI, ANIELLE SALVIANO DE ALMEIDA; HENRIQUE, PEDRO ANTONIO FRANCA; VALDIVIESO, DANIELA; SANCHEZ-LIMACHE, DANIEL ENRIQUE; SILVA, GABRIELA ROBERTO; et al. Insertion of a Divergent GAF-like Domain Defines a Novel Family of YcgR Homologues That Bind c-di-GMP in Leptospirales. ACS OMEGA, v. 10, n. 4, p. 19-pg., 2025-01-21. (20/04680-0, 19/00195-2, 21/05262-0, 21/00070-5, 21/10577-0, 17/17303-7, 23/10432-7, 17/06394-1, 16/09047-8, 23/18211-0, 22/08730-7)

DE ALMEIDA, MICHELLE TEIXEIRA; BARBOSA, ANA PAULA; BOMFIM, CAMILA GASQUE; VISNARDI, ALINE BIAZOLA; VINCES, TANIA CHURASACARI; CERONI, ALEXANDRE; DURIGON, EDISON LUIZ; GUZZO, CRISTIANE RODRIGUES. Obtaining a high titer of polyclonal antibodies from rats to the SARS-CoV-2 nucleocapsid protein and its N- and C-terminal domains for diagnostic test development. JOURNAL OF IMMUNOLOGICAL METHODS, v. 522, p. 9-pg., 2023-09-27. (21/10577-0, 19/21739-0, 22/08730-7, 20/14914-8, 19/00195-2, 16/09047-8, 20/04680-0)

AMORIM, VITOR MARTINS DE FREITAS; SOARES, EDUARDO PEREIRA; FERRARI, ANIELLE SALVIANO DE ALMEIDA; MERIGHI, DAVI GABRIEL SALUSTIANO; DE SOUZA, ROBSON FRANCISCO; GUZZO, CRISTIANE RODRIGUES; DE SOUZA, ANACLETO SILVA. 3-Chymotrypsin-like Protease (3CLpro) of SARS-CoV-2: Validation as a Molecular Target, Proposal of a Novel Catalytic Mechanism, and Inhibitors in Preclinical and Clinical Trials. Viruses-Basel, v. 16, n. 6, p. 22-pg., 2024-06-01. (21/10577-0, 22/08730-7, 19/00195-2, 21/00070-5, 23/13894-1, 16/09047-8, 20/04680-0)

DE ALMEIDA, MICHELLE TEIXEIRA; MERIGHI, DAVI GABRIEL SALUSTIANO; VISNARDI, ALINE BIAZOLA; GONCALVES, CAUE AUGUSTO BONETO; AMORIM, VITOR MARTINS DE FREITAS; FERRARI, ANIELLE SALVIANO DE ALMEIDA; DE SOUZA, ANACLETO SILVA; GUZZO, CRISTIANE RODRIGUES. Latin America's Dengue Outbreak Poses a Global Health Threat. Viruses-Basel, v. 17, n. 1, p. 32-pg., 2025-01-01. (23/13894-1, 21/05262-0, 21/10577-0, 23/18211-0, 22/08730-7, 23/07366-2)