Isolation and structural studies on the Xanthomonas citri Type IV pilus and T4SS pilus

• Grant number: 19/12234-2
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: July 01, 2019
• End date: June 30, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Shaker Chuck Farah
• Grantee: Edgar Enrique Llontop Cornejo
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 17/17303-7 - Structure and function of bacterial secretion systems, AP.TEM

Abstract

The pili of T2SS, T3SS, T4SS and T4P mediate the attachment of bacteria to other structures which can be other cells, phages or inanimate surfaces. This attachment can often lead to the transfer of virulence factors, invasion by a phage or the formation of biofilms. Type IV pili are specialized versions type II secretion systems (T2SSs) that translocate folded proteins from the periplasm across the cellular envelope of Gram-negative bacteria. The secreted pilin subunits form an extracellular helical polymer that can extend to several times the length of the bacterial cell. Associated proteins (adhesins, minor pilins) often facilitate T4P attachment to surfaces. Pilus polymerization, attachment and retraction (termed twitching motility) allow bacteria to move across surfaces and orient themselves with respect to other bacteria, and so play a very important role in the formation of the highly organized and multicellular structures in bacterial biofilms. We have been able to identify conditions in which the X. citri T4P are expressed, as well as a protocol for their purification. We recently been able to visiualize T4P on the surface of X. citri cells by electron microscopy. We have also studied several of regulatory proteins responsible for pilus biogenesis. We therefore plan to undertake a project to determine the molecular structure of coponents of the X. citri T4P by X-ray crystallography and Cryo-EM. Finally, we have recently sequenced the genome of the bacteriophage Xcm4-11 that infects X. citri via attachement to its Type IV pilus. We threfore plan to determine the structure of this baceriophage by Cryo-EM.

Scientific publications

(4)

(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)

VENTURA FERNANDES, BIANCA H.; FEITOSA, NATALIA MARTINS; BARBOSA, ANA PAULA; BOMFIM, CAMILA GASQUE; GARNIQUE, ANALI M. B.; ROSA, IVANA F.; RODRIGUES, MAIRA S.; DORETTO, LUCAS B.; COSTA, DANIEL F.; CAMARGO-DOS-SANTOS, BRUNO; et al. oxicity of spike fragments SARS-CoV-2 S protein for zebrafish: A tool to study its hazardous for human health. Science of The Total Environment, v. 813, MAR 20 2022. (19/00195-2, 18/07098-0, 20/05761-3, 17/17303-7, 20/04680-0, 14/04294-1, 19/21739-0, 19/19939-1, 19/18356-2, 19/14285-3, 19/12234-2)

CENENS, WILLIAM; ANDRADE, MAXUEL O.; LLONTOP, EDGAR; ALVAREZ-MARTINEZ, CRISTINA E.; SGRO, GERMAN G.; FARAH, CHUCK S.. Bactericidal type IV secretion system homeostasis in Xanthomonas citri. PLOS PATHOGENS, v. 16, n. 5, MAY 2020. (19/12234-2, 15/18237-2, 14/04294-1, 17/17303-7, 11/07777-5)

LLONTOP, EDGAR E.; CENENS, WILLIAM; FAVARO, DENIZE C.; SGRO, GERMAN G.; SALINAS, ROBERTO K.; GUZZO, CRISTIANE R.; FARAH, CHUCK S.. he PilB-PilZ-FimX regulatory complex of the Type IV pilus from Xanthomonas citr. PLOS PATHOGENS, v. 17, n. 8, AUG 2021. (15/18237-2, 19/12234-2, 17/17303-7)

HESPANHOL, JULIA TAKUNO; SANCHEZ-LIMACHE, DANIEL ENRIQUE; NICASTRO, GIANLUCCA GONCALVES; MEAD, LIAM; LLONTOP, EDGAR ENRIQUE; CHAGAS-SANTOS, GUSTAVO; FARAH, CHUCK SHAKER; DE SOUZA, ROBSON FRANCISCO; GALHARDO, RODRIGO DA SILVA; LOVERING, ANDREW L.; et al. Antibacterial T6SS effectors with a VRR-Nuc domain are structure-specific nucleases. eLIFE, v. 11, p. 26-pg., 2022-10-13. (17/17303-7, 19/12234-2, 21/03400-6, 16/09047-8, 20/15389-4, 18/25316-4, 18/04553-8, 17/02178-2, 19/22715-8)