Activation of a yeast replication origin near a double-stranded DNA break.

Abstract:

:Irradiation in the G1 phase of the cell cycle delays the onset of DNA synthesis and transiently inhibits the activation of replication origins in mammalian cells. It has been suggested that this inhibition is the result of the loss of torsional tension in the DNA after it has been damaged. Because irradiation causes DNA damage at an undefined number of nonspecific sites in the genome, it is not known how cells respond to limited DNA damage, and how replication origins in the immediate vicinity of a damage site would behave. Using the sequence-specific HO endonuclease, we have created a defined double-stranded DNA break in a centromeric plasmid in G1-arrested cells of the yeast Saccharomyces cerevisiae. We show that replication does initiate at the origin on the cut plasmid, and that the plasmid replicates early in the S phase after linearization in vivo. These observations suggest that relaxation of a supercoiled DNA domain in yeast need not inactivate replication origins within that domain. Furthermore, these observations rule out the possibility that the late replication context associated with chromosomal termini is a consequence of DNA ends.

journal_name

Genes Dev

journal_title

Genes & development

authors

Raghuraman MK,Brewer BJ,Fangman WL

doi

10.1101/gad.8.5.554

subject

Has Abstract

pub_date

1994-03-01 00:00:00

pages

554-62

issue

5

eissn

0890-9369

issn

1549-5477

journal_volume

8

pub_type

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