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Functional dynamics of Orc1b protein during the life cycle of Trypanosoma

Grant number: 17/07693-2
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2018
Effective date (End): November 21, 2021
Field of knowledge:Biological Sciences - Parasitology
Principal researcher:Maria Carolina Quartim Barbosa Elias Sabbaga
Grantee:Marcela de Oliveira Vitarelli
Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Associated research grant:13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling, AP.CEPID
Associated scholarship(s):19/04483-2 - Functional dynamics of Orc1b protein: a ChIP-seq analysis, BE.EP.DR

Abstract

Trypanosma cruzi is a protozoan which belongs to the Trypanosomatidae family and it is the etiological agent of Chagas Disease, a neglected disease that affects millions of people worldwide. This parasite alternates between replicative (epi and amastigotes) and non-replicative (trypomastigotes) forms in its hosts. Therefore it needs intense control of DNA replication, which involves coordinated action of several proteins. DNA replication starts with the assembly of the pre-replication complex (pre-RC) component ORC onto replication origins, located near the nucleosomes. Followed by the recruitment of other proteins from pre-RC, including Orc1b, whose function is not yet clear. Preliminary data of Marques, et al. (2016) indicates Orc1b participation as a positive regulator in the replication instead of having only a structural role in the ORC complex. Preliminary data from our group suggest the interaction of Orc1b with histones presenting specific post-translational modifications. Thus, we aim to deepen the studies of the Orc1b role in the DNA replication of T. cruzi and its possible interactors. Assays will be performed to analyze the expression of this protein in the course of T. cruzi cell and life cycle, and also the production of hemi knockouts to access the impact of the absence of this protein in the replication. In addition, immunoprecipitation, pull down, ChIP-seq and ELISA assays will be executed in order to analyze the possible interactors of Orc1b, with emphasis on the modified histones H4K10ac and H3K76Me3. This knowledge will support the development of future antiparasitic strategies and it will also be useful to further understand the biology of these parasites. (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)
DA SILVA, MARCELO S.; VITARELLI, MARCELA O.; SOUZA, BRUNO F.; ELIAS, MARIA CAROLINA. Comparative Analysis of the Minimum Number of Replication Origins in Trypanosomatids and Yeasts. GENES, v. 11, n. 5 MAY 2020. Web of Science Citations: 0.
DA SILVA, MARCELO S.; CAYRES-SILVA, GUSTAVO R.; VITARELLI, MARCELA O.; MARIN, PAULA A.; HIRAIWA, PRISCILA M.; ARAUJO, CHRISTIANE B.; SCHOLL, BRUNO B.; AVILA, ANDREA R.; MCCULLOCH, RICHARD; REIS, MARCELO S.; ELIAS, MARIA CAROLINA. Transcription activity contributes to the firing of non-constitutive origins in African trypanosomes helping to maintain robustness in S-phase duration. SCIENTIFIC REPORTS, v. 9, DEC 6 2019. Web of Science Citations: 0.

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