Mechanisms of DNA damage recognition and processing in chromatin by DNA polymerase lambda
Active Project Grant Awarded Last action
- Award ID
- F31ES039118
- Recipient
-
University Of Kansas Medical Center Research Institute, Inc. UEI YXJGGNC5J269
- Awarding agency
-
National Institute of Environmental Health Sciences
National Institutes of Health, Department of Health and Human Services
- Obligated
- $78,020
Award facts
- Assistance listing
- 93.113 Environmental Health
- Completion date 0% complete
- Originally
- Place of performance
- Kansas City, KS 66160, USA
Award details come from USAspending.gov.
Description
- Abstract
- Humans are regularly exposed to environmental toxins including air pollutants, heavy metals, and cigarette smoke, which generate reactive oxygen species (ROS) capable of damaging genomic DNA. These exposures cause thousands of DNA lesions including mutagenic oxidative base lesions (e.g., 8OXOG) or single-strand breaks (ssbs). Ssbs are among the most frequent DNA lesions and if not repaired can drive genome instability and contribute to environmentally induced human diseases. In cells, ssbs arise both directly from ROS attack to the DNA backbone and indirectly during base excision repair (BER) as DNA glycosylases and AP- endonuclease 1 process oxidative base lesions. Both BER and SSB repair converge to a common DNA intermediate containing a one nucleotide (1-NT GAP) that requires GAP filling DNA synthesis by DNA polymerase beta (POL β) or DNA polymerase lambda (POL λ). Importantly, ROS induce DNA damage ubiquitously throughout the genome and therefore repair mechanisms must contend with the structural constraints of genomic DNA that is packaged into chromatin through the repeating unit known as the nucleosome. Prior studies show POL β engages nucleosomal 1-NT gaps and performs GAP filling DNA synthesis in vitro, though with a lower efficiency compared to non-nucleosomal DNA. In comparison, the mechanism of POL λ-mediated DNA repair within the nucleosome is completely unknown. The overarching goal of this proposal is to define how POL λ searches for, recognizes, and fills 1-NT gaps in the nucleosome and to understand how these mechanisms protect cells from environmentally induced oxidative stress. To address this, i propose the following aims: (1) determine the mechanism POL λ uses to perform GAP filling DNA synthesis in the nucleosome, (2) define the DNA damage search and recognition mechanisms of POL λ, and (3) determine the contribution of POL λ to oxidative DNA damage repair in chromatin within cells. To complete these aims, i will integrate a combination of biochemical, structural, and cellular biology techniques to comprehensively study the mechanisms used by POL λ during 1-NT GAP repair within chromatin. Specifically, cryo-electron microscopy, binding assays, and enzyme kinetics will be used to gain insight into the mechanism used by POL λ to perform GAP filling DNA synthesis in the nucleosome (aim 1). Correlative optical tweezers and fluorescence microscopy (ctfm) will be used to determine the mechanisms POL λ uses to search for DNA damage in non-nucleosomal and nucleosomal DNA (aim 2). Finally, cellular fractionation and comet assays will be used to characterize the role of POL λ in cellular repair of oxidative DNA damage (aim 3). These advanced techniques will provide me with significant technical training under the supervision of an exceptional sponsorship team. In addition to technical skills, the training plan will support my scientific development through improving scientific communication and mentorship skills to provide me with a strong foundation for a successful career in biomedical research.
Transactions
| Action | Obligated | Date |
|---|---|---|
| Amendment | $39,010 | |
| Amendment | $39,010 | |
| Original award | $0 |
Quick answers
Who received award F31ES039118?
F31ES039118 was awarded to University Of Kansas Medical Center Research Institute, Inc. by National Institute of Environmental Health Sciences (National Institutes of Health) on January 13, 2027.
How much is award F31ES039118 worth?
$78,020 has been obligated on F31ES039118.
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