Mechanistic of the phosphate-uptake and regulation: the close relationship between an ATP-binding cassette transporter and a two-component system

• Grant number: 20/10171-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: January 01, 2021
• End date: August 31, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Andrea Balan Fernandes
• Grantee: Lucas Souza dos Santos
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

ABC transporters are membrane proteins involved in a wide range of biological activities such as cellular metabolism and signaling, adhesion, biofilm formation, and pathogenesis, in both prokaryotic and eukaryotic cells. Among these, the PstSCABPhoU, together with the two-component system PhoR/PhoB, participates in the transport of phosphate and other pathogenesis factors in Xanthomonas citri, the causal agent of the citrus canker. This phytopathology spreads through commercial citrus crops, generating great losses in agricultural production in Brazil and the rest of the world. There is no effective control for the disease, hence the development of new molecular strategies of inhibition of the canker is urgent and essential. In our laboratory, we have achieved good progress in the production, purification, and characterization of components of this complex. However, the mechanisms on how the interaction between these components of PstSCAB-PhoU-PhoR/PhoB remains unclear. Thus, we aim, by application of X-ray crystallography and single-particle cryo-EM, to solve the structures of the complex and perform interaction assay between PstSCAB-PhoU, PhoU-PhoR, and PhoR-PhoB using techniques such as Circular Dichroism (CD), Small-Angle X-Ray Scattering (SAXS), Isothermal Titration Calorimetry (ITC), Microscale Thermophoresis (MST), and pull-down assays. The project is highly feasible and will benefit from the expertise of the LBEA-ICB/USP team together with Prof. Dr. Albert Guskov from the University of Groningen (the Netherlands). We also have the support of Prof. Dr. Cristiano Oliveira - Instituto de Física/USP and the infrastructure of CNPEM through LNLS, LNBio, and LNNano. Altogether, we expect to achieve consistent results that are going to be impactful for the development of new strategies of control for the citrus canker, and other related pathologies. (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)

DOS SANTOS, NAARA M.; PICINATO, BEATRIZ A.; SANTOS, LUCAS S.; DE ARAUJO, HUGO L.; BALAN, ANDREA; KOIDE, TIE; MARQUES, MARILIS V.. Mapping the IscR regulon sheds light on the regulation of iron homeostasis in Caulobacter. FRONTIERS IN MICROBIOLOGY, v. 15, p. 18-pg., 2024-09-30. (21/10577-0, 22/00308-4, 20/10171-0, 22/06360-8, 18/17309-8)