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Rap1 Binds Single-stranded DNA at Telomeric Double- and Single-stranded Junctions and Competes with Cdc13 Protein

Gustafsson, Cecilia LU ; Rhodin, Jenny LU and Cohn, Marita LU (2011) In Journal of Biological Chemistry 286(52). p.45174-45185
Abstract
The ends of eukaryotic chromosomes are protected by specialized telomere chromatin structures. Rap1 and Cdc13 are essential for the formation of functional telomere chromatin in budding yeast by binding to the double-stranded part and the single-stranded 3' overhang, respectively. We analyzed the binding properties of Saccharomyces castellii Rap1 and Cdc13 to partially single-stranded oligonucleotides, mimicking the junction of the double- and single-stranded DNA (ds-ss junction) at telomeres. We determined the optimal and the minimal DNA setup for a simultaneous binding of Rap1 and Cdc13 at the ds-ss junction. Remarkably, Rap1 is able to bind to a partially single-stranded binding site spanning the ds-ss junction. The binding over the... (More)
The ends of eukaryotic chromosomes are protected by specialized telomere chromatin structures. Rap1 and Cdc13 are essential for the formation of functional telomere chromatin in budding yeast by binding to the double-stranded part and the single-stranded 3' overhang, respectively. We analyzed the binding properties of Saccharomyces castellii Rap1 and Cdc13 to partially single-stranded oligonucleotides, mimicking the junction of the double- and single-stranded DNA (ds-ss junction) at telomeres. We determined the optimal and the minimal DNA setup for a simultaneous binding of Rap1 and Cdc13 at the ds-ss junction. Remarkably, Rap1 is able to bind to a partially single-stranded binding site spanning the ds-ss junction. The binding over the ds-ss junction is anchored in a single double-stranded hemi-site and is stabilized by a sequence-independent interaction of Rap1 with the single-stranded 3' overhang. Thus, Rap1 is able to switch between a sequence-specific and a nonspecific binding mode of one hemi-site. At a ds-ss junction configuration where the two binding sites partially overlap, Rap1 and Cdc13 are competing for the binding. These results shed light on the end protection mechanisms and suggest that Rap1 and Cdc13 act together to ensure the protection of both the 3' and the 5' DNA ends at telomeres. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Rap1, Saccharomyces castellii, Cdc13, 3'overhang, Telomeres, Telomerase, Molecular genetics, Molecular cell biology, DNA-protein interaction, Naumovozyma castellii
in
Journal of Biological Chemistry
volume
286
issue
52
pages
45174 - 45185
publisher
ASBMB
external identifiers
  • wos:000298645500083
  • pmid:22075002
  • scopus:84455173182
ISSN
1083-351X
DOI
10.1074/jbc.M111.300517
language
English
LU publication?
yes
id
a4b2703e-f7d4-4301-bfbd-522e237699cd (old id 2303928)
date added to LUP
2012-01-31 11:25:50
date last changed
2017-09-24 03:05:45
@article{a4b2703e-f7d4-4301-bfbd-522e237699cd,
  abstract     = {The ends of eukaryotic chromosomes are protected by specialized telomere chromatin structures. Rap1 and Cdc13 are essential for the formation of functional telomere chromatin in budding yeast by binding to the double-stranded part and the single-stranded 3' overhang, respectively. We analyzed the binding properties of Saccharomyces castellii Rap1 and Cdc13 to partially single-stranded oligonucleotides, mimicking the junction of the double- and single-stranded DNA (ds-ss junction) at telomeres. We determined the optimal and the minimal DNA setup for a simultaneous binding of Rap1 and Cdc13 at the ds-ss junction. Remarkably, Rap1 is able to bind to a partially single-stranded binding site spanning the ds-ss junction. The binding over the ds-ss junction is anchored in a single double-stranded hemi-site and is stabilized by a sequence-independent interaction of Rap1 with the single-stranded 3' overhang. Thus, Rap1 is able to switch between a sequence-specific and a nonspecific binding mode of one hemi-site. At a ds-ss junction configuration where the two binding sites partially overlap, Rap1 and Cdc13 are competing for the binding. These results shed light on the end protection mechanisms and suggest that Rap1 and Cdc13 act together to ensure the protection of both the 3' and the 5' DNA ends at telomeres.},
  author       = {Gustafsson, Cecilia and Rhodin, Jenny and Cohn, Marita},
  issn         = {1083-351X},
  keyword      = {Rap1,Saccharomyces castellii,Cdc13,3'overhang,Telomeres,Telomerase,Molecular genetics,Molecular cell biology,DNA-protein interaction,Naumovozyma castellii},
  language     = {eng},
  number       = {52},
  pages        = {45174--45185},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Rap1 Binds Single-stranded DNA at Telomeric Double- and Single-stranded Junctions and Competes with Cdc13 Protein},
  url          = {http://dx.doi.org/10.1074/jbc.M111.300517},
  volume       = {286},
  year         = {2011},
}