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Characterization of LmRad1 of Leishmania major and study of its role in the DNA damage response

Grant number: 13/26806-1
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): August 01, 2014
Effective date (End): September 30, 2017
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Luiz Ricardo Orsini Tosi
Grantee:Elaine Vieira Santos
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:13/50219-9 - Studying the control of gene expression in Leishmania: post-translational modification, non coding RNAs, cis-elements and gene amplification, AP.TEM


The protozoan parasite Leishmania has a plastic and dynamic genome where gene amplification and chromosome translocation are relatively common phenomena. The molecular basis for these processes are likely to generate DNA structures that resemble damaged DNA. In higher eukaryotes, the cellular checkpoint control system detects these structures and blocks cell cycle progression, allowing for the required DNA repair. In mammalian and yeast cells, the trimeric complex 9-1-1 (Rad9, Rad1 and Hus1) participates in the initial steps of recognition and signaling of replicative stress. We have recently published the characterization of Leishmania major LmHus1 and LmRad9 proteins. We showed structural and phylogenetic similarities between these proteins and those from the 9-1-1 complex from other eukaryotes. The aim of this project is to investigate the existence of the missing Rad1 homologue in L. major. To do this, we propose to investigate the expression of a candidate homologue and study its role in the DNA damage response of the parasite. We will generate cell lines overexpressing LmRad1, as well as LmRad1 knockout parasites. The subcellular localization of the protein and its interaction with LmHus1 and LmRad9 will also be investigated. (AU)

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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)
DAMASCENO, JEZIEL D.; OBONAGA, RICARDO; SANTOS, ELAINE V.; SCOTT, ALAN; MCCULLOCH, RICHARD; TOSI, LUIZ R. O.. Functional compartmentalization of Rad9 and Hus1 reveals diverse assembly of the 9-1-1 complex components during the DNA damage response in Leishmania. Molecular Microbiology, v. 101, n. 6, p. 1054-1068, . (13/00570-1, 14/00751-9, 13/26806-1, 14/06824-8, 09/54014-7)
SANTOS, RENATO E. R. S.; SILVA, GABRIEL L. A.; SANTOS, ELAINE V.; DUNCAN, SAMUEL M.; MOTTRAM, JEREMY C.; DAMASCENO, JEZIEL D.; TOSI, LUIZ R. O.. A DiCre recombinase-based system for inducible expression in Leishmania major. Molecular and Biochemical Parasitology, v. 216, p. 45-48, . (14/00751-9, 13/26806-1, 14/06824-8, 13/00570-1, 09/54014-7)

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