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Investigation of the structural requirements for recognition of T4SS-related toxins by the coupling protein VirD4.

Grant number: 18/09277-9
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Effective date (Start): June 01, 2018
Effective date (End): July 31, 2022
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal Investigator:Shaker Chuck Farah
Grantee:Gabriel Umaji Oka
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

VirD4 is an ATPase that selects proteins to be secreted in most T4SSs. We have shown that VirD4 from Xac recognizes conserved domains called XVIPCD found at the C-termini of X-Tfes (Alegria et al., 2005). We are interested in understanding the structure of the VirD4-XVIPCD complex, how the complex is transfered to the rest of the T4SS machinery and the nature of the role played by ATP binding and/or hydrolysis. Since the transfer of toxins by the X. citri T4SS kills recipient E. coli cells, we will produce a X. citri strain in which all of the genes coding for X-Tfes (XVIPS) have been knocked out. This non-killer strain, with an otherwise functional T4SS, will be transformed with plasmids expressing specific X-Tfes. If this is successful, this assay could be used to quantitatively measure transfer modified X-Tfes with mutations in specific recognition motifs. This X. citri strain will also be used to screen mutations in other T4SS components to map the structural requirements for secretion.

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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)
OKA, GABRIEL U.; SOUZA, DIORGE P.; CENENS, WILLIAM; MATSUYAMA, BRUNO Y.; CARDOSO, MARCUS V. C.; OLIVEIRA, LUCIANA C.; LIMA, FILIPE DA SILVA; CUCCOVIA, IOLANDA M.; GUZZO, CRISTIANE R.; SALINAS, ROBERTO K.; et al. tructural basis for effector recognition by an antibacterial type IV secretion syste. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v. 119, n. 1, . (18/09277-9, 15/18237-2, 17/17303-7, 16/00458-5)

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