Telomerase is a specialized reverse transcriptase (RT) that synthesizes telomere DNA repeats at chromosome ends, using only a very short region of its intrinsic telomerase RNA (TR) subunit as template. This highly specialized function of telomerase relies on a special mechanism whereby the template RNA and the telomeric DNA dissociate and realign during the processive synthesis of repeats. However, the detailed mechanism of telomerase template translocation remains to be determined. This research program aims to articulate the unique mechanism of telomerase action and identify elements that regulate specific steps of template translocation. Although telomerase uses the single-stranded telomeric DNA as its native substrate, we have recently discovered that telomerase can act as a conventional RT utilizing RNA/DNA duplex as substrate. More surprisingly, telomerase recognizes the duplex substrate with a sequence-specificity. These crucial findings have provided great insights into the molecular mechanism of telomerase action. We hypothesize that duplex- binding and duplex-dissociation are important steps of the telomere-repeat synthesis cycle, and regulate telomere-repeat addition rate and processivity.
Specific Aims of the research program include (1) Determining the role duplex-binding affinity in template translocation efficiency, (2) Determining the rate-limiting step of template translocation, and (3) characterize the sequence- dependent termination of nucleotide addition by telomerase. We expect the outcomes of these experiments will greatly add to our understanding of telomerase mechanism.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM094450-02
Application #
8307315
Study Section
Molecular Genetics A Study Section (MGA)
Program Officer
Gerratana, Barbara
Project Start
2011-08-01
Project End
2016-05-31
Budget Start
2012-06-01
Budget End
2013-05-31
Support Year
2
Fiscal Year
2012
Total Cost
$283,322
Indirect Cost
$93,322
Name
Arizona State University-Tempe Campus
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
943360412
City
Tempe
State
AZ
Country
United States
Zip Code
85287
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Podlevsky, Joshua D; Chen, Julian J-L (2016) Evolutionary perspectives of telomerase RNA structure and function. RNA Biol 13:720-32
Podlevsky, Joshua D; Li, Yang; Chen, Julian J-L (2016) The functional requirement of two structural domains within telomerase RNA emerged early in eukaryotes. Nucleic Acids Res 44:9891-9901
Qi, Xiaodong; Rand, Dustin P; Podlevsky, Joshua D et al. (2015) Prevalent and distinct spliceosomal 3'-end processing mechanisms for fungal telomerase RNA. Nat Commun 6:6105
Huang, Jing; Bley, Christopher J; Rand, Dustin P et al. (2015) Sample Preparation of Telomerase Subunits for Crystallization. Bio Protoc 5:
Stanley, Susan E; Chen, Julian J L; Podlevsky, Joshua D et al. (2015) Telomerase mutations in smokers with severe emphysema. J Clin Invest 125:563-70
Huang, Jing; Brown, Andrew F; Wu, Jian et al. (2014) Structural basis for protein-RNA recognition in telomerase. Nat Struct Mol Biol 21:507-12
Brown, Andrew F; Podlevsky, Joshua D; Qi, Xiaodong et al. (2014) A self-regulating template in human telomerase. Proc Natl Acad Sci U S A 111:11311-6

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