Functional characterization of CCNA_02417 and CCNA_02418 genes during DNA repair in Caulobacter crescentus

• Grant number: 22/04993-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2022
• End date: December 31, 2023
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Rodrigo da Silva Galhardo
• Grantee: Henrique Kustor Antonini
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 19/19435-3 - The role of DNA damage and mitochondrial function in vascular, immune and neurological ageing (DNA MoVINg), AP.TEM

Abstract

Genetic sequencing of Caulobacter crescentus allowed the identification of open reading frames (ORFs) and genes in its genome. Some of these genes are conserved, making it possible to predict their function. On the other hand, others have not yet been adequately characterized, including the operon formed by the genes CCNA_02417 and CCNA_02418, which encodes DNA repair proteins that, together, are believed to configure a repair system never described before, part of the SOS response in C. crescentus. Unpublished data from our group indicate that both genes are involved in DNA repair, configuring a possible new light-independent photoproduct repair system that acts on non-sporulating vegetative cells. It was proposed that CCNA_02417 and CCNA_02418 have complementary activity in the repair of uracil dimers, formed by the deamination of cytosine dimers, which are photo products so that the correction of this DNA lesion will be effective and will avoid mutations only in the presence of both of them. Thus, these proteins are expected to have physical proximity during their activity. CCNA_02418 has a domain of the unknown function (DUF 4130) that is often found in operons containing genes encoding proteins similar to CCNA_02417. As a consequence of this project, we intend to confirm the interaction between CCNA_02417 and CCNA_02418 through the two-hybrid system, specify the domain of the second that participates in the interaction, if it occurs, and also check this operon's messenger RNA stability in mutants CCNA_02417 and CCNA_02418 via qRT-PCR to correlate the results with complementation assays already concluded by our research group.(AU)