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M200322200v1
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Papers In Press, published online ahead of print February 12, 2002
J. Biol. Chem, 10.1074/jbc.M200322200
Submitted on January 11, 2002
Revised on February 12, 2002
Accepted on February 11, 2002

Scheduled conversion of replication complex architecture at replication origins of S. cerevisiae during the cell cycle

Ryusuke Tadokoro, Masako Fujita, Hitoshi Miura, Katsuhiko Shirahige, Hiroshi Yoshikawa, Toshiki Tsurimoto, and Chikashi Obuse

Biological Science, Nara Institute of Science and Technology, Ikoma, Nara 630-0101

Corresponding Author: c-obuse{at}bs.aist-nara.ac.jp

Replication of DNA within S. cerevisiae chromosomes is initiated from multiple origins, whose activation follow their own inherent time schedules during the S phase of the cell cycle. It has been demonstrated that a characteristic replicative complex (RC) that includes ORC (origin recognition complex) is formed at each origin and shifts between post- and pre-replicative states during the cell cycle. We wanted to determine if there was an association between this shift in the state of the RC and firing events at replication origins. Time course analyses of RC architecture using UV-footprinting with synchronously growing cells revealed that pre-replicative states at both early and late firing origins appeared simultaneously during late M phase, remained in this state during G1 phase, and converted to the post-replicative state at various times during S phase. Since the conversion of the origin footprinting profiles and origin firing, as assessed by 2D-gel electrophoresis, occurred concomitantly at each origin, then these two events must be closely related. However, conversion of the late firing origin occurred without actual firing. This was observed when the late origin was suppressed in clb5 deficient cells and a replication fork originating from an outside origin replicated the late origin passively. This mechanism ensures that replication at each chromosomal locus occurs only once per cell cycle by shifting existing pre-RCs to the post-RC state, when it is replicated without firing.


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This article has been cited by other articles:


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J. Biol. Chem.Home page
S. Ohta, Y. Tatsumi, M. Fujita, T. Tsurimoto, and C. Obuse
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[Abstract] [Full Text] [PDF]




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