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Novel components of Homologous Recombination Repair in the parasite Toxoplasma gondii

Award Number: R01AI129807
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 04/07/2017
PERIOD OF PERFORMANCE END DATE: 03/31/2022

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Novel components of Homologous Recombination Repair in the parasite Toxoplasma gondii - The protozoan parasite Toxoplasma gondii is an important human and veterinary pathogen. In humans, acute disease is caused by rapidly growing `tachyzoites' stage. In this context, we hypothesize that during this intense tachyzoite propagation, replication-associated DNA stress could appear to be generating fork collapse and DNA double strand breaks which should be repaired by homologous recombination repair (HRR). The hypothesis is supported by the presence of basal γH2A.X, a marker of DSB, in the tachyzoite as well as the inhibition of its replication by the KU55933, an inhibitor of ATM, a key kinase in the HRR pathway. In addition, our analysis indicates that key molecules seem to be not present in HRR pathway in T. gondii. Differences in the HRR cascade imply an opportunity for the identification of novel and specific HRR components in T. gondii. If available, such factors could represent attractive candidates for the development of new drugs against Toxoplasmosis. Our goal is to identify novel components of HRR and analyze their importance in HRR pathway. In specific aim 1 we will develop a methodology named iPOND to pull down components of HRR in T. gondii and their identification and analysis by bioinformatics tools. In addition, we will analyze their biological significance in tachyzoite replication and DSB sensitivity, together with some HRR components yet identified by in silico analysis (Mre11, RAD51, ATM, BRCA2, BRCT domain containing proteins). In specific aim 2 we will design a model of HR in vivo by generating tachyzoites conditionally expressing the endonuclease I-SceI to generate DSB specifically in a plasmid incorporated by transfection. This plasmid contains two versions of GFP inactive genes in which the functional gene is expressed only after HR.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2023 ( Subtotal = $0 )
2023	2021	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445	SAN MARTIN	BUENOS AIRES			ARG	Allergy and Infectious Diseases Research	000	5	4/20/2023	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2021 ( Subtotal = $126,131 )
2021	2021	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES			ARG	Allergy and Infectious Diseases Research	000	5	4/16/2021	NON-COMPETING CONTINUATION	$126,131
														Subtotal = $126,131

Issue Date FY: 2020 ( Subtotal = $127,942 )
2020	2020	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	000	4	4/9/2020	NON-COMPETING CONTINUATION	$127,942
														Subtotal = $127,942

Issue Date FY: 2019 ( Subtotal = $129,700 )
2019	2019	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	002	3	4/3/2019	NON-COMPETING CONTINUATION	$129,700
2019	2018	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	001	2	4/3/2019	NON-COMPETING CONTINUATION	$0
2019	2017	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	000	1	4/3/2019	NEW	$0
														Subtotal = $129,700

Issue Date FY: 2018 ( Subtotal = $133,248 )
2018	2018	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	000	2	3/27/2018	NON-COMPETING CONTINUATION	$133,248
														Subtotal = $133,248

Issue Date FY: 2017 ( Subtotal = $132,795 )
2017	2017	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	001	1	7/28/2017	NEW	-$132,795
2017	2017	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	002	1	7/31/2017	NEW	$132,795
2017	2017	FUNDACION INSTITUTO DE INVESTIGACIONES BIOTECNOLOGICAS	COLECTORA DE AVENIDA GENERAL PAZ 5445, EDIFICIO 24, CC 30	SAN MARTIN	BUENOS AIRES	B1650KNA		ARG	Allergy and Infectious Diseases Research	000	1	4/5/2017	NEW	$132,795
														Subtotal = $132,795

Grand Total All Awards = $649,816

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Novel components of Homologous Recombination Repair in the parasite Toxoplasma gondii

Award Number: R01AI129807

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 04/07/2017

PERIOD OF PERFORMANCE END DATE: 03/31/2022

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