Uncoupling global and fine-tuning replication timing determinants for mouse pericentric heterochromatin

Rong Wu, Prim B Singh, David M Gilbert

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Mouse chromocenters are clusters of late-replicating pericentric heterochromatin containing HP1 bound to trimethylated lysine 9 of histone H3 (Me3K9H3). Using a cell-free system to initiate replication within G1-phase nuclei, we demonstrate that chromocenters acquire the property of late replication coincident with their reorganization after mitosis and the establishment of a global replication timing program. HP1 dissociated during mitosis but rebound before the establishment of late replication, and removing HP1 from chromocenters by competition with Me3K9H3 peptides did not result in early replication, demonstrating that this interaction is neither necessary nor sufficient for late replication. However, in cells lacking the Suv39h1,2 methyltransferases responsible for K9H3 trimethylation and HP1 binding at chromocenters, replication of chromocenter DNA was advanced by 10-15% of the length of S phase. Reintroduction of Suv39h1 activity restored the later replication time. We conclude that Suv39 activity is required for the fine-tuning of pericentric heterochromatin replication relative to other late-replicating domains, whereas separate factors establish a global replication timing program during early G1 phase.

Original languageEnglish
Pages (from-to)185-94
Number of pages10
JournalJournal of Cell Biology
Volume174
Issue number2
DOIs
Publication statusPublished - Jul 17 2006

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Heterochromatin
G1 Phase
Mitosis
Cell-Free System
DNA Replication
S Phase
Histones
Lysine
Peptides
methylcobalamin-coenzyme M methyltransferase

Keywords

  • Animals
  • Cells, Cultured
  • Centrosome
  • Chromosomal Proteins, Non-Histone
  • DNA Replication Timing
  • DNA, Satellite
  • Fibroblasts
  • G1 Phase
  • Heterochromatin
  • Histones
  • Lysine
  • Methylation
  • Methyltransferases
  • Mice
  • Protein Binding
  • Repressor Proteins
  • Xenopus
  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

Cite this

Uncoupling global and fine-tuning replication timing determinants for mouse pericentric heterochromatin. / Wu, Rong; Singh, Prim B; Gilbert, David M.

In: Journal of Cell Biology, Vol. 174, No. 2, 17.07.2006, p. 185-94.

Research output: Contribution to journalArticle

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abstract = "Mouse chromocenters are clusters of late-replicating pericentric heterochromatin containing HP1 bound to trimethylated lysine 9 of histone H3 (Me3K9H3). Using a cell-free system to initiate replication within G1-phase nuclei, we demonstrate that chromocenters acquire the property of late replication coincident with their reorganization after mitosis and the establishment of a global replication timing program. HP1 dissociated during mitosis but rebound before the establishment of late replication, and removing HP1 from chromocenters by competition with Me3K9H3 peptides did not result in early replication, demonstrating that this interaction is neither necessary nor sufficient for late replication. However, in cells lacking the Suv39h1,2 methyltransferases responsible for K9H3 trimethylation and HP1 binding at chromocenters, replication of chromocenter DNA was advanced by 10-15{\%} of the length of S phase. Reintroduction of Suv39h1 activity restored the later replication time. We conclude that Suv39 activity is required for the fine-tuning of pericentric heterochromatin replication relative to other late-replicating domains, whereas separate factors establish a global replication timing program during early G1 phase.",
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AB - Mouse chromocenters are clusters of late-replicating pericentric heterochromatin containing HP1 bound to trimethylated lysine 9 of histone H3 (Me3K9H3). Using a cell-free system to initiate replication within G1-phase nuclei, we demonstrate that chromocenters acquire the property of late replication coincident with their reorganization after mitosis and the establishment of a global replication timing program. HP1 dissociated during mitosis but rebound before the establishment of late replication, and removing HP1 from chromocenters by competition with Me3K9H3 peptides did not result in early replication, demonstrating that this interaction is neither necessary nor sufficient for late replication. However, in cells lacking the Suv39h1,2 methyltransferases responsible for K9H3 trimethylation and HP1 binding at chromocenters, replication of chromocenter DNA was advanced by 10-15% of the length of S phase. Reintroduction of Suv39h1 activity restored the later replication time. We conclude that Suv39 activity is required for the fine-tuning of pericentric heterochromatin replication relative to other late-replicating domains, whereas separate factors establish a global replication timing program during early G1 phase.

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