Extended DNA binding site in Pot1 broadens sequence specificity to allow recognition of heterogeneous fission yeast telomeres

Kelly M. Trujillo, Jeremy T. Bunch, Peter Baumann

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The Pot1 (protection of telomeres) protein binds to single-stranded telomeric DNA and is essential for the protection of chromosome ends from degradation and end-to-end fusions. The Pot1 amino-terminal DNA binding domain, Pot1N, adopts an oligonucleotide/oligosaccharide binding fold and binds GGTTAC motifs cooperatively and with exceptionally high sequence specificity. We have now examined DNA binding to naturally occurring telomeric substrates based on the analysis of 100 cloned chromosome ends and in the context of the full-length Pot1 protein. Here, we describe several important differences between Pot1 and Pot1N with apparent consequences for chromosome end protection. Specifically, full-length Pot1-DNA complexes are more stable, and the minimal binding site for a Pot1 monomer is extended into two adjacent telomeric repeats. We provide evidence that Pot1 contains a second DNA binding motif that recognizes DNA with reduced sequence specificity compared with the domain present in Pot1N. The two DNA binding motifs cooperate, whereby the amino-terminal oligonucleotide/ oligosaccharide binding fold determines the registry of binding, and the internal DNA binding motif stabilizes the complex and expands the protected region toward the 3′-end. Consistent with a role in chromosome end capping, Pot1 prevents access of telomerase to the 3′-end and protects against exonucleolytic degradation.

Original languageEnglish (US)
Pages (from-to)9119-9128
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number10
DOIs
StatePublished - Mar 11 2005

Fingerprint

Schizosaccharomyces
Telomere
Yeast
Nucleotide Motifs
Chromosomes
Binding Sites
DNA
Oligosaccharides
Oligonucleotides
Single-Stranded DNA
Telomerase
Registries
Proteins
Degradation
Fusion reactions
Monomers
Substrates

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Extended DNA binding site in Pot1 broadens sequence specificity to allow recognition of heterogeneous fission yeast telomeres. / Trujillo, Kelly M.; Bunch, Jeremy T.; Baumann, Peter.

In: Journal of Biological Chemistry, Vol. 280, No. 10, 11.03.2005, p. 9119-9128.

Research output: Contribution to journalArticle

Trujillo, Kelly M. ; Bunch, Jeremy T. ; Baumann, Peter. / Extended DNA binding site in Pot1 broadens sequence specificity to allow recognition of heterogeneous fission yeast telomeres. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 10. pp. 9119-9128.
@article{7090d94ef911467fabac92632ce664ef,
title = "Extended DNA binding site in Pot1 broadens sequence specificity to allow recognition of heterogeneous fission yeast telomeres",
abstract = "The Pot1 (protection of telomeres) protein binds to single-stranded telomeric DNA and is essential for the protection of chromosome ends from degradation and end-to-end fusions. The Pot1 amino-terminal DNA binding domain, Pot1N, adopts an oligonucleotide/oligosaccharide binding fold and binds GGTTAC motifs cooperatively and with exceptionally high sequence specificity. We have now examined DNA binding to naturally occurring telomeric substrates based on the analysis of 100 cloned chromosome ends and in the context of the full-length Pot1 protein. Here, we describe several important differences between Pot1 and Pot1N with apparent consequences for chromosome end protection. Specifically, full-length Pot1-DNA complexes are more stable, and the minimal binding site for a Pot1 monomer is extended into two adjacent telomeric repeats. We provide evidence that Pot1 contains a second DNA binding motif that recognizes DNA with reduced sequence specificity compared with the domain present in Pot1N. The two DNA binding motifs cooperate, whereby the amino-terminal oligonucleotide/ oligosaccharide binding fold determines the registry of binding, and the internal DNA binding motif stabilizes the complex and expands the protected region toward the 3′-end. Consistent with a role in chromosome end capping, Pot1 prevents access of telomerase to the 3′-end and protects against exonucleolytic degradation.",
author = "Trujillo, {Kelly M.} and Bunch, {Jeremy T.} and Peter Baumann",
year = "2005",
month = "3",
day = "11",
doi = "10.1074/jbc.M414511200",
language = "English (US)",
volume = "280",
pages = "9119--9128",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "10",

}

TY - JOUR

T1 - Extended DNA binding site in Pot1 broadens sequence specificity to allow recognition of heterogeneous fission yeast telomeres

AU - Trujillo, Kelly M.

AU - Bunch, Jeremy T.

AU - Baumann, Peter

PY - 2005/3/11

Y1 - 2005/3/11

N2 - The Pot1 (protection of telomeres) protein binds to single-stranded telomeric DNA and is essential for the protection of chromosome ends from degradation and end-to-end fusions. The Pot1 amino-terminal DNA binding domain, Pot1N, adopts an oligonucleotide/oligosaccharide binding fold and binds GGTTAC motifs cooperatively and with exceptionally high sequence specificity. We have now examined DNA binding to naturally occurring telomeric substrates based on the analysis of 100 cloned chromosome ends and in the context of the full-length Pot1 protein. Here, we describe several important differences between Pot1 and Pot1N with apparent consequences for chromosome end protection. Specifically, full-length Pot1-DNA complexes are more stable, and the minimal binding site for a Pot1 monomer is extended into two adjacent telomeric repeats. We provide evidence that Pot1 contains a second DNA binding motif that recognizes DNA with reduced sequence specificity compared with the domain present in Pot1N. The two DNA binding motifs cooperate, whereby the amino-terminal oligonucleotide/ oligosaccharide binding fold determines the registry of binding, and the internal DNA binding motif stabilizes the complex and expands the protected region toward the 3′-end. Consistent with a role in chromosome end capping, Pot1 prevents access of telomerase to the 3′-end and protects against exonucleolytic degradation.

AB - The Pot1 (protection of telomeres) protein binds to single-stranded telomeric DNA and is essential for the protection of chromosome ends from degradation and end-to-end fusions. The Pot1 amino-terminal DNA binding domain, Pot1N, adopts an oligonucleotide/oligosaccharide binding fold and binds GGTTAC motifs cooperatively and with exceptionally high sequence specificity. We have now examined DNA binding to naturally occurring telomeric substrates based on the analysis of 100 cloned chromosome ends and in the context of the full-length Pot1 protein. Here, we describe several important differences between Pot1 and Pot1N with apparent consequences for chromosome end protection. Specifically, full-length Pot1-DNA complexes are more stable, and the minimal binding site for a Pot1 monomer is extended into two adjacent telomeric repeats. We provide evidence that Pot1 contains a second DNA binding motif that recognizes DNA with reduced sequence specificity compared with the domain present in Pot1N. The two DNA binding motifs cooperate, whereby the amino-terminal oligonucleotide/ oligosaccharide binding fold determines the registry of binding, and the internal DNA binding motif stabilizes the complex and expands the protected region toward the 3′-end. Consistent with a role in chromosome end capping, Pot1 prevents access of telomerase to the 3′-end and protects against exonucleolytic degradation.

UR - http://www.scopus.com/inward/record.url?scp=15744372316&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=15744372316&partnerID=8YFLogxK

U2 - 10.1074/jbc.M414511200

DO - 10.1074/jbc.M414511200

M3 - Article

C2 - 15637058

AN - SCOPUS:15744372316

VL - 280

SP - 9119

EP - 9128

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 10

ER -