Replication factory activation can be decoupled from the replication timing program by modulating Cdk levels. / Thomson, Alexander M.; Gillespie, Peter J.; Blow, J. Julian.
In: Journal of Cell Biology, Vol. 188, No. 2, 25.01.2010, p. 209-221.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Replication factory activation can be decoupled from the replication timing program by modulating Cdk levels
A1 - Thomson,Alexander M.
A1 - Gillespie,Peter J.
A1 - Blow,J. Julian
AU - Thomson,Alexander M.
AU - Gillespie,Peter J.
AU - Blow,J. Julian
PY - 2010/1/25
Y1 - 2010/1/25
N2 - <p>In the metazoan replication timing program, clusters of replication origins located in different subchromosomal domains fire at different times during S phase. We have used Xenopus laevis egg extracts to drive an accelerated replication timing program in mammalian nuclei. Although replicative stress caused checkpoint-induced slowing of the timing program, inhibition of checkpoint kinases in an unperturbed S phase did not accelerate it. Lowering cyclin-dependent kinase (Cdk) activity slowed both replication rate and progression through the timing program, whereas raising Cdk activity increased them. Surprisingly, modest alteration of Cdk activity changed the amount of DNA synthesized during different stages of the timing program. This was associated with a change in the number of active replication factories, whereas the distribution of origins within active factories remained relatively normal. The ability of Cdks to differentially effect replication initiation, factory activation, and progression through the timing program provides new insights into the way that chromosomal DNA replication is organized during S phase.</p>
AB - <p>In the metazoan replication timing program, clusters of replication origins located in different subchromosomal domains fire at different times during S phase. We have used Xenopus laevis egg extracts to drive an accelerated replication timing program in mammalian nuclei. Although replicative stress caused checkpoint-induced slowing of the timing program, inhibition of checkpoint kinases in an unperturbed S phase did not accelerate it. Lowering cyclin-dependent kinase (Cdk) activity slowed both replication rate and progression through the timing program, whereas raising Cdk activity increased them. Surprisingly, modest alteration of Cdk activity changed the amount of DNA synthesized during different stages of the timing program. This was associated with a change in the number of active replication factories, whereas the distribution of origins within active factories remained relatively normal. The ability of Cdks to differentially effect replication initiation, factory activation, and progression through the timing program provides new insights into the way that chromosomal DNA replication is organized during S phase.</p>
KW - CYCLIN-DEPENDENT KINASES
KW - XENOPUS EGG EXTRACTS
KW - S-PHASE CHECKPOINT
KW - DNA-REPLICATION
KW - CELL-CYCLE
KW - DORMANT ORIGINS
KW - BUDDING YEAST
KW - FISSION YEAST
KW - CHROMOSOMAL DOMAINS
KW - REPLICON CLUSTERS
U2 - 10.1083/jcb.200911037
DO - 10.1083/jcb.200911037
M1 - Article
JO - Journal of Cell Biology
JF - Journal of Cell Biology
SN - 0021-9525
IS - 2
VL - 188
SP - 209
EP - 221
ER -