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DNA helicases and associated factors in genome stability

Award Number: R35GM133437
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/01/2019
PERIOD OF PERFORMANCE END DATE: 07/31/2024

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DNA helicases and associated factors in genome stability - PROJECT SUMMARY DNA helicases function in virtually all aspects of DNA replication, recombination, and repair. As such, they are vital to maintaining genome integrity and are disease linked when mutated. Despite many in vivo and in vitro advances in working with helicases, there is a gap in knowledge connecting mutant alleles of helicase genes to the treatment of patients in clinics. The objective of my research is to gain mechanistic insight into how DNA helicases function in genome maintenance and why their dysfunction leads to disease. Toward this goal, we are studying PIF1 and RecQ family helicases, which are evolutionarily conserved and because mutations in the human genes encoding these helicases are associated with multiple diseases. Our current work focuses on the roles of RecQ helicases in DNA inter-strand crosslink (ICL) repair and RecQ and Pif1 helicases in telomere maintenance. To perform this work, we will employ a variety of classic and cutting edge experimental techniques, from standard in vitro enzymatic assays and model organism genetics to next- generation sequencing, crosslinking mass spectrometry, and the development of custom click chemistry probes. Overall, this work will provide fundamental data critical to understanding how PIF1 and RecQ family helicases aid in the maintenance of genome stability, and it will ultimately lead to therapeutic targets and treatments for helicase-linked diseases. I am requesting funds to purchase a Refeyn TwoMP mass photometer. This device enables rapid mass measurements of small volumes of dilute biomolecules, with single-molecule precision. We routinely generate recombinant protein preparations for biochemical assays, and knowing the oligomeric state and monodispersity under physiological conditions is critical to the design, analysis, and interpretation of our experiments. We currently use size-exclusion chromatography (SEC) to determine recombinant protein mass, but SEC is slow, has low resolution, and requires large volumes of very concentrated protein. Therefore, SEC limits the availability of our chromatography system and is incompatible with proteins that cannot be purified in large amounts or do not tolerate the concentration process. The purchase of a mass photometer will provide a rapid and high-resolution solution to these problems with a dedicated device that is inexpensive and easy to use, improving our biophysical capabilities and supporting all of our projects.


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 = $642,554 )
2023	2023	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	003	5	9/4/2023	SUPPLEMENT FOR EXPANSION	$222,360
2023	2023	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	002	5	8/10/2023	SUPPLEMENT FOR EXPANSION	$24,079
2023	2023	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	001	5	6/30/2023	NON-COMPETING CONTINUATION	$387,694
2023	2023	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	000	4	5/27/2023	SUPPLEMENT FOR EXPANSION	$8,421
														Subtotal = $642,554

Issue Date FY: 2022 ( Subtotal = $522,027 )
2022	2022	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	002	4	8/17/2022	SUPPLEMENT FOR EXPANSION	$72,232
2022	2022	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	003	4	9/12/2022	SUPPLEMENT FOR EXPANSION	$13,540
2022	2022	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	001	4	8/11/2022	NON-COMPETING CONTINUATION	$388,102
2022	2022	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	000	3	12/9/2021	SUPPLEMENT FOR EXPANSION	$48,153
														Subtotal = $522,027

Issue Date FY: 2021 ( Subtotal = $439,868 )
2021	2021	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	001	3	9/17/2021	SUPPLEMENT FOR EXPANSION	$59,139
2021	2021	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	000	3	7/7/2021	NON-COMPETING CONTINUATION	$380,729
														Subtotal = $439,868

Issue Date FY: 2020 ( Subtotal = $377,773 )
2020	2020	TRUSTEES OF INDIANA UNIVERSITY	107 S INDIANA AVENUE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	000	2	7/24/2020	NON-COMPETING CONTINUATION	$377,773
														Subtotal = $377,773

Issue Date FY: 2019 ( Subtotal = $372,813 )
2019	2019	TRUSTEES OF INDIANA UNIVERSITY	BRYAN HALL 107 S INDIANA AVENUE	BLOOMINGTON	IN	47405	MONROE	USA	Biomedical Research and Research Training	000	1	7/26/2019	NEW	$372,813
														Subtotal = $372,813

Grand Total All Awards = $2,355,035

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DNA helicases and associated factors in genome stability

Award Number: R35GM133437

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 08/01/2019

PERIOD OF PERFORMANCE END DATE: 07/31/2024

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