TRF2 recruits ORC through TRFH domain dimerization.

Publisher: Elsevier Country of Publication: Netherlands NLM ID: 101731731 Publication Model: Print-Electronic Cited Medium: Print ISSN: 0167-4889 (Print) Linking ISSN: 01674889 NLM ISO Abbreviation: Biochim Biophys Acta Mol Cell Res Subsets: MEDLINE

Original Publication: Amsterdam : Elsevier

Chromatin/*chemistry ; Minichromosome Maintenance Proteins/*metabolism ; Origin Recognition Complex/*metabolism ; Telomere/*metabolism ; Telomeric Repeat Binding Protein 2/*metabolism; Cell Line, Tumor ; Chromatin/metabolism ; DNA Replication ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Gene Expression ; HEK293 Cells ; HeLa Cells ; Humans ; Lac Repressors/genetics ; Lac Repressors/metabolism ; Minichromosome Maintenance Proteins/genetics ; Mutation ; Origin Recognition Complex/genetics ; Osteoblasts/cytology ; Osteoblasts/metabolism ; Protein Domains ; Protein Multimerization ; Signal Transduction ; Telomere/ultrastructure ; Telomeric Repeat Binding Protein 1/genetics ; Telomeric Repeat Binding Protein 1/metabolism ; Telomeric Repeat Binding Protein 2/chemistry ; Telomeric Repeat Binding Protein 2/genetics

Telomeres are specialized chromatin structures that prevent the degradation and instability of the ends of linear chromosomes. While telomerase maintains long stretches of the telomeric repeat, the majority of telomeric DNA is duplicated by conventional DNA replication. A fundamental step in eukaryotic DNA replication involves chromatin binding of the origin recognition complex (ORC). In human cells, telomeric repeat binding factor 2 (TRF2) is thought to play a role in the recruitment of ORC onto telomeres. To better understand the mechanism of TRF2-mediated ORC recruitment, we utilized a lacO-LacI protein tethering system in U2OS cells and found that ectopically targeted TRF2, but not TRF1, can recruit ORC onto the lacO array. We further found that the TRF homology (TRFH) dimerization domain of TRF2, but not its mutant defective in dimerization, is sufficient for ORC and minichromosome maintenance (MCM) recruitment. Mutations impairing the dimerization also compromised ORC recruitment by full-length TRF2. Similar results were obtained using immunoprecipitation and GST pull-down assays. Together, these results suggest that dimerized TRF2 recruits ORC and stimulates pre-replication complex (pre-RC) formation at telomeres through the TRFH domain.
(Copyright © 2016 Elsevier B.V. All rights reserved.)

Keywords: DNA replication; MCM; ORC; Pre-replication complex; TRF2; Telomere

0 (Chromatin)
0 (Escherichia coli Proteins)
0 (Lac Repressors)
0 (LacI protein, E coli)
0 (Origin Recognition Complex)
0 (TERF2 protein, human)
0 (Telomeric Repeat Binding Protein 1)
0 (Telomeric Repeat Binding Protein 2)
EC 3.6.4.12 (Minichromosome Maintenance Proteins)

Date Created: 20161113 Date Completed: 20171004 Latest Revision: 20180919

20221213

10.1016/j.bbamcr.2016.11.004

27836746