CryoEM study of the adsorption of a phage to a type IV pilus

• Grant number: 23/13894-1
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: March 01, 2024
• End date: April 30, 2027
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Cristiane Rodrigues Guzzo Carvalho
• Grantee: Davi Gabriel Salustiano Merighi
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 21/10577-0 - Biology of Bacteria and Bacteriophages Research Center, AP.CEPID

Abstract

As causative agent of citrus canker, Xanthomonas citri is an economically revelant phytopathogen threatening Brazilian and global agriculture. The ability to communicate using autoinducer molecules, through Quorum Sensing, makes it possible to modulate gene expression in a coordinated manner and initiate the production of various virulence factors, such as proteases and endoglucanase, in addition to having a repertoire of specialized machinery that assists in colonization of the host and establishment of infection, including secretion systems, secretion of outer membrane vesicles and extracellular appendages such as the type IV pilus (T4P). Despite conferring an advantage to the bacteria by functioning as an adsorption mechanism and mediating twitching motility, on the other hand, T4P can serve as a binding site for bacteriophages that use it as an entry mechanism to initiate infection. ¦Xacm4-11, for example, is a podovirus that requires the functional T4P extension and retraction system to complete its life cycle. In the constant battle between bacteria and bacteriophages, many defense and attack mechanisms on both sides are developed, such as CRISPR-Cas/9 or Gabija defense systems. Continuing the projects developed in the first part of the doctorate, we intend to study bacterial communication by determining the structure of its Quorum Sensing molecule belonging to the DSF family by analyzing previously obtained Nuclear Magnetic Resonance results. Furthermore, we also propose to solve the structure of the ¦Xacm4-11 phage tail in complex with T4P by Cryo-EM to unravel the mechanisms of the interaction in the primordial stage of infection, in addition to investigating the nature and determining the structure of the protein complexes with co-purified with phages, evaluating the possibility of their participation as a defense mechanism against infection.

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)

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)

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)