See instaicKons);
We aim to understand how telomeres solve the end-protection problem. Specifically, this proposal describesexperiments to gain insight Into the mechanism by which the telomeric protein complex, shelterin, repressesthe two main DNA repair reactions, NHEJ and HDR at chromosome ends.
Our aim I s to delineate the natureof the repair reactions at telomeres, their outcomes, what factors are involved, how these pathways areregulated, and how telomeres prevent inappropriate pnscessing by repair pathways.Telomere dysfunction is thought to be main source of genome instability in human cancer, has beenimplicated in aging, and is at the core of several human diseases that are caused by loss of telomeric DNA(e.g. dyskeratosis congenita, anaplastic anemia). When telomeres become dysfunctional, inappropriaterepair at chromosome ends is a major determinant of the pathological outcomes. Detailed understanding ofthe repair pathways at dysfunctional telomeres is therefore expected to impact the treatment and preventionhuman disease.We will use genetic, molecular genetic, and cell biological techniques to address the mechanism by whichtelomeres repress inappropriate repair and define the pathological repair reactions at dysfunctionaltelomeres. The experimental approaches will involve the tools for studying mammalian telomeres that wehave developed under the auspices of this grant over the past 16 years. Specifically, we will use mouseembryo fibroblast cell lines from which shelterin components can be deleted with the Cre recombinase. Bychanging the genetic background of these cells, using compound genotypes, we can monitor the repair atdysfuncUonanl telomere, delineate the genes involved In the reactions, and define how shelterin repressesthese reactions at functional telomeres.

Public Health Relevance

;A growing number of human disease states are now understood to involve a failure in the function oftelomeres, the protective elements at the ends of human chromosomes. This project is aimed at gainingInsight into how telomeres protect chromosome ends and what happens when this protection is lost. Thework is expected to be significant in the context of cancer, aging, and telomere-related diseases such asdyskeratosis congetica.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37GM049046-18
Application #
7784291
Study Section
Special Emphasis Panel (NSS)
Program Officer
Carter, Anthony D
Project Start
1993-05-01
Project End
2015-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
18
Fiscal Year
2010
Total Cost
$541,836
Indirect Cost
Name
Rockefeller University
Department
Anatomy/Cell Biology
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Kibe, Tatsuya; Zimmermann, Michal; de Lange, Titia (2016) TPP1 Blocks an ATR-Mediated Resection Mechanism at Telomeres. Mol Cell 61:236-46
Frescas, David; de Lange, Titia (2014) TRF2-tethered TIN2 can mediate telomere protection by TPP1/POT1. Mol Cell Biol 34:1349-62
Kabir, Shaheen; Hockemeyer, Dirk; de Lange, Titia (2014) TALEN gene knockouts reveal no requirement for the conserved human shelterin protein Rap1 in telomere protection and length regulation. Cell Rep 9:1273-80
Frescas, David; de Lange, Titia (2014) A TIN2 dyskeratosis congenita mutation causes telomerase-independent telomere shortening in mice. Genes Dev 28:153-66
Zimmermann, Michal; Kibe, Tatsuya; Kabir, Shaheen et al. (2014) TRF1 negotiates TTAGGG repeat-associated replication problems by recruiting the BLM helicase and the TPP1/POT1 repressor of ATR signaling. Genes Dev 28:2477-91
Frescas, David; de Lange, Titia (2014) Binding of TPP1 protein to TIN2 protein is required for POT1a,b protein-mediated telomere protection. J Biol Chem 289:24180-7
Zimmermann, Michal; de Lange, Titia (2014) 53BP1: pro choice in DNA repair. Trends Cell Biol 24:108-17
Doksani, Ylli; Wu, John Y; de Lange, Titia et al. (2013) Super-resolution fluorescence imaging of telomeres reveals TRF2-dependent T-loop formation. Cell 155:345-356
Zimmermann, Michal; Lottersberger, Francisca; Buonomo, Sara B et al. (2013) 53BP1 regulates DSB repair using Rif1 to control 5' end resection. Science 339:700-4
Ahmed, Emad A; Sfeir, Agnel; Takai, Hiroyuki et al. (2013) Ku70 and non-homologous end joining protect testicular cells from DNA damage. J Cell Sci 126:3095-104

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