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PMID:21852327

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Citation

Choi, KH, Farrell, AS, Lakamp, AS and Ouellette, MM (2011) Characterization of the DNA binding specificity of Shelterin complexes. Nucleic Acids Res. 39:9206-23

Abstract

The Shelterin complex associates with telomeres and plays an essential role in telomere protection and telomerase regulation. In its most abundant form, the complex is composed of six core components: TRF1, TRF2, POT1, TIN2, TPP1 and RAP1. Of these subunits, three can interact directly with either single-stranded (POT1) or double-stranded (TRF1, TRF2) telomeric DNA. In this report, we have developed assays to measure the DNA binding activity of Shelterin complexes in human cell extracts. With these assays, we have characterized the composition and DNA binding specificity of two Shelterin complexes: a 6-member complex that contains all six core components and a second complex that lacks TRF1. Our results show that both of these complexes bind with high affinity (K(D) = 1.3-1.5 × 10(-9) M) and selectively to ds/ss-DNA junctions that carry both a binding site for POT1 (ss-TTAGGGTTAG) and a binding site for the SANT/Myb domain of TRF1 or TRF2 (ds-TTAGGGTTA). This DNA binding specificity suggests the preferential recruitment of these complexes to areas of the telomere where ss- and ds-DNA are in close proximity, such as the 3'-telomeric overhang, telomeric DNA bubbles and the D-loop at the base of T-loops.

Links

PubMed PMC3241663 Online version:10.1093/nar/gkr665

Keywords

Cell Line; DNA/metabolism; HeLa Cells; Humans; Telomere/metabolism; Telomere-Binding Proteins/metabolism; Telomeric Repeat Binding Protein 2/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:TERF1

enables

GO:0003691: double-stranded telomeric DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:TERF1

part_of

GO:0070187: shelterin complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ACD

part_of

GO:0070187: shelterin complex

ECO:0000315: mutant phenotype evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:TE2IP

part_of

GO:0070187: shelterin complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:TERF2

enables

GO:0003691: double-stranded telomeric DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:TERF2

part_of

GO:0070187: shelterin complex

ECO:0000315: mutant phenotype evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:TINF2

part_of

GO:0070187: shelterin complex

ECO:0000315: mutant phenotype evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:POTE1

enables

GO:0098505: G-rich strand telomeric DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:POTE1

part_of

GO:0070187: shelterin complex

ECO:0000315: mutant phenotype evidence used in manual assertion

C

Seeded From UniProt

complete

Notes

See also

References

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