Cloning and optmization of the expression conditions of LIC_11299 to LIC_11302 genic cluster

• Grant number: 18/08996-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2018
• End date: July 31, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Cristiane Rodrigues Guzzo Carvalho
• Grantee: Nathalia Marins de Azevedo
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Leptospirosis is an infectious disease of global importance and distribution, it is caused by the Gram negative spirochete Leptospira interrogans. The contaminated person may present suddenly high fever, myalgia, headache, malaise, meningitis, and may also cause major liver and kidney problems that can lead to the patient's death. In most cases the disease development is benign and an improvement in the clinical condition is observed in about seven days. The project here presented is focused on collaborate with the understanding of the signaling pathway of the bacterial second messenger, c-di-GMP, that mediates the bacterial way of life and regulates different phenotypes, such like: biofilms, motility, genic transcription among others. The protein to be studied in this project is the LIC_11300 that presents five transmembrane helix in the N-terminal area along with a GGDEF motif, believed to be active, in the C-terminal area of the protein. The analysis of the LIC_11300 genic context showed that the proteins LIC_11299 to LIC_11302 form a cluster and might be involved in the same signaling pathway. Interestingly, the LIC_11301 is also a membrane protein and presents a transmembrane helix in its N-terminal area, along with a germane motif in its C-terminal section. The LIC_11302 belongs to the Haloacid dehalogenase family of proteins, and could act as a pyrophosphatase. The LIC_11299 gene is pointed to the opposite direction of the LIC_11300 gene in the L. interrogans gene, the fact that it is a putative lipoprotein made it interesting. In that way, our hypothesis is that the LIC_11299 to LIC_11302 genes might participate in the same signaling pathway, mediated by the second messenger, c-di-GMP. The project will be given by the interaction study between the proteins through cloning, expression and analysis of the results obtained.

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)

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)