Involvement of human ORC and TRF2 in pre-replication complex assembly at telomeres

Genes Cells. 2008 Oct;13(10):1045-59. doi: 10.1111/j.1365-2443.2008.01224.x. Epub 2008 Aug 29.

Abstract

The origin recognition complex (ORC) binds to replication origins to regulate the cell cycle-dependent assembly of pre-replication complexes (pre-RCs). We have found a novel link between pre-RC assembly regulation and telomere homeostasis in human cells. Biochemical analyses showed that human ORC binds to TRF2, a telomere sequence-binding protein that protects telomeres and functions in telomere length homeostasis, via the ORC1 subunit. Immunostaining further revealed that ORC and TRF2 partially co-localize in nuclei, whereas chromatin immunoprecipitation analyses confirmed that pre-RCs are assembled at telomeres in a cell cycle-dependent manner. Over-expression of TRF2 stimulated ORC and MCM binding to chromatin and RNAi-directed TRF2 silencing resulted in reduced ORC binding and pre-RC assembly at telomeres. As expected from previous reports, TRF2 silencing induced telomere elongation. Interestingly, ORC1 silencing by RNAi weakened the TRF2 binding as well as the pre-RC assembly at telomeres, suggesting that ORC and TRF2 interact with each other to achieve stable binding. Furthermore, ORC1 silencing also resulted in modest telomere elongation. These data suggest that ORC might be involved in telomere homeostasis in human cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle / physiology
  • Cell Line, Transformed
  • Cell Nucleus / metabolism
  • Chromatin Immunoprecipitation
  • DNA Replication*
  • Fibroblasts / metabolism
  • HeLa Cells / metabolism
  • Humans
  • Origin Recognition Complex / metabolism*
  • Telomere / genetics
  • Telomere / metabolism*
  • Telomeric Repeat Binding Protein 2 / metabolism*

Substances

  • Origin Recognition Complex
  • TERF2 protein, human
  • Telomeric Repeat Binding Protein 2