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Multiple DNA Interactions Contribute to the Initiation of Telomerase Elongation

Karademir Andersson, Ahu LU ; Gustafsson, Cecilia LU ; Krishnankutty, Roopesh LU and Cohn, Marita LU (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
organization
publishing date
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
  • scopus:85020065973
  • wos:000404702800001
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
2017-09-18 11:37:00
@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},
  keyword      = {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},
  volume       = {429},
  year         = {2017},
}