Structure of the Xanthomonas citri type IV pilus secretin

• Grant number: 18/21076-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: May 01, 2019
• End date: July 31, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Cristiane Rodrigues Guzzo Carvalho
• Grantee: Gabriel Guarany de Araujo
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 17/17303-7 - Structure and function of bacterial secretion systems, AP.TEM
• Associated scholarship(s): 19/26988-9 - Cryo-EM study of the adsorption of a type IV pilus-specific bacteriophage to its receptor, BE.EP.DR

Abstract

Type IV pili (T4P) are long retracting extracellular bacterial filaments that allow motility and orientation over surfaces. In Xanthomonas species, a genus with notorious phytopathogenic members, T4P were associated with biofilm formation, adhesion, and virulence processes. The Xanthomonas T4P secretion machinery is a large protein complex that crosses the cell's membranes, by which pilus subunits are polymerized and translocated to the external medium. This PhD project aims to study the secretin subcomplex that forms the membrane channel that the filament goes through. For this purpose, its detailed three-dimensional structure will be determined with cryo-electron microscopy (cryo-EM). The secretin is composed of PilQ, which multimerizes to form the channel. This protein will be cloned into vectors for the addition of purification tags and expressed, in parallel, in vivo (in Xanthomonas citri pv citrus 306) and heterologously in E. coli. Its corresponding gene will be deleted and complemented with plasmids, and it will be checked if the in vivo phenotypic properties of the edited T4P machinery were thus maintained. The secretin will be purified from these cultures and characterized by SEC-MALS and transmission electron microscopy before being taken to cryo-EM for structural determination. Finally, it is expected to obtain a model of this complex essential for T4P that will guide future studies in Xanthomonas. (AU)

Scientific publications

(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)

DANIELLE BASTOS ARAUJO; RAFAEL RAHAL GUARAGNA MACHADO; DEYVID EMANUEL AMGARTEN; FERNANDA DE MELLO MALTA; GABRIEL GUARANY DE ARAUJO; CAIRO OLIVEIRA MONTEIRO; ERIKA DONIZETTI CANDIDO; CAMILA PEREIRA SOARES; FERNANDO GATTI DE MENEZES; ANA CAROLINA CORNACHIONI PIRES; et al. SARS-CoV-2 isolation from the first reported patients in Brazil and establishment of a coordinated task network. Memórias do Instituto Oswaldo Cruz, v. 115, 2020-10-23. (16/20045-7, 17/25123-9, 18/21076-9, 17/24769-2, 18/23680-0, 20/04680-0, 20/06409-1)

BECERRA, JOHANA; ARAUJO, GABRIEL G.; VASQUEZ-PONCE, FELIPE; LAMADRID-FERIS, FARIDE; LINCOPAN, NILTON. ACT-107, a novel variant of AmpC β-lactamase from Enterobacter huaxiensis isolated from Neotropical leaf frog (Phyllomedusa distincta) inhabiting the Brazilian Atlantic Forest. JOURNAL OF GLOBAL ANTIMICROBIAL RESISTANCE, v. 33, p. 7-pg., 2023-06-08. (20/08224-9, 14/16320-7, 18/21076-9)

ALINE DIAS DA, PURIFICACAO; NATHALIA MARINS DE, AZEVEDO; GABRIEL GUARANY DE, ARAUJO; ROBSON FRANCISCO DE, SOUZA; RODRIGUES, GUZZO CRISTIANE. The World of Cyclic Dinucleotides in Bacterial Behavior. Molecules, v. 25, n. 10, MAY 2020. (18/21076-9, 16/08414-7, 17/10611-8, 18/08996-1, 19/00195-2)