Multiple DNA Interactions Contribute to the Initiation of Telomerase Elongation
(2017) In Journal of Molecular Biology 429(14). p.2109-2123- Abstract
Telomerase maintains telomere length and chromosome integrity by adding short tandem repeats of single-stranded DNA to the 3' ends, via reverse transcription of a defined template region of its RNA subunit. To further understand the telomerase elongation mechanism, we studied the primer utilization and extension activity of the telomerase from the budding yeast Naumovozyma castellii (Saccharomyces castellii), which displays a processive nucleotide and repeat addition polymerization. For the efficient initiation of canonical elongation, telomerase required 4-nt primer 3' end complementarity to the template RNA. This DNA-RNA hybrid formation was highly important for the stabilization of an initiation-competent telomerase-DNA complex.... (More)
Telomerase maintains telomere length and chromosome integrity by adding short tandem repeats of single-stranded DNA to the 3' ends, via reverse transcription of a defined template region of its RNA subunit. To further understand the telomerase elongation mechanism, we studied the primer utilization and extension activity of the telomerase from the budding yeast Naumovozyma castellii (Saccharomyces castellii), which displays a processive nucleotide and repeat addition polymerization. For the efficient initiation of canonical elongation, telomerase required 4-nt primer 3' end complementarity to the template RNA. This DNA-RNA hybrid formation was highly important for the stabilization of an initiation-competent telomerase-DNA complex. Anchor site interactions with the DNA provided additional stabilization to the complex. Our studies indicate three additional separate interactions along the length of the DNA primer, each providing different and distinct contributions to the initiation event. A sequence-independent anchor site interaction acts immediately adjacent to the base-pairing 3' end, indicating a protein anchor site positioned very close to the catalytic site. Two additional anchor regions further 5' on the DNA provide sequence-specific contributions to the initiation of elongation. Remarkably, a non-telomeric sequence in the distal 25- to 32-nt region negatively influences the initiation of telomerase elongation, suggesting an anchor site with a regulatory role in the telomerase elongation decision.
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- author
- Karademir Andersson, Ahu LU ; Gustafsson, Cecilia LU ; Krishnankutty, Roopesh LU and Cohn, Marita LU
- organization
- publishing date
- 2017-07-07
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- DNA-protein interaction, Naumovozyma castellii, Repeat addition processivity, Telomerase substrate specificity, Telomere
- in
- Journal of Molecular Biology
- volume
- 429
- issue
- 14
- pages
- 15 pages
- publisher
- Elsevier
- external identifiers
-
- pmid:28506636
- wos:000404702800001
- scopus:85020065973
- ISSN
- 0022-2836
- DOI
- 10.1016/j.jmb.2017.04.023
- language
- English
- LU publication?
- yes
- id
- 7516c5b3-252f-43ff-94de-5e8abe7e2f4a
- date added to LUP
- 2017-06-29 10:49:56
- date last changed
- 2024-02-29 17:36:19
@article{7516c5b3-252f-43ff-94de-5e8abe7e2f4a,
abstract = {{<p>Telomerase maintains telomere length and chromosome integrity by adding short tandem repeats of single-stranded DNA to the 3' ends, via reverse transcription of a defined template region of its RNA subunit. To further understand the telomerase elongation mechanism, we studied the primer utilization and extension activity of the telomerase from the budding yeast Naumovozyma castellii (Saccharomyces castellii), which displays a processive nucleotide and repeat addition polymerization. For the efficient initiation of canonical elongation, telomerase required 4-nt primer 3' end complementarity to the template RNA. This DNA-RNA hybrid formation was highly important for the stabilization of an initiation-competent telomerase-DNA complex. Anchor site interactions with the DNA provided additional stabilization to the complex. Our studies indicate three additional separate interactions along the length of the DNA primer, each providing different and distinct contributions to the initiation event. A sequence-independent anchor site interaction acts immediately adjacent to the base-pairing 3' end, indicating a protein anchor site positioned very close to the catalytic site. Two additional anchor regions further 5' on the DNA provide sequence-specific contributions to the initiation of elongation. Remarkably, a non-telomeric sequence in the distal 25- to 32-nt region negatively influences the initiation of telomerase elongation, suggesting an anchor site with a regulatory role in the telomerase elongation decision.</p>}},
author = {{Karademir Andersson, Ahu and Gustafsson, Cecilia and Krishnankutty, Roopesh and Cohn, Marita}},
issn = {{0022-2836}},
keywords = {{DNA-protein interaction; Naumovozyma castellii; Repeat addition processivity; Telomerase substrate specificity; Telomere}},
language = {{eng}},
month = {{07}},
number = {{14}},
pages = {{2109--2123}},
publisher = {{Elsevier}},
series = {{Journal of Molecular Biology}},
title = {{Multiple DNA Interactions Contribute to the Initiation of Telomerase Elongation}},
url = {{http://dx.doi.org/10.1016/j.jmb.2017.04.023}},
doi = {{10.1016/j.jmb.2017.04.023}},
volume = {{429}},
year = {{2017}},
}