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

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

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Replication of DNA within Saccharomyces 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 an origin recognition complex is formed at each origin and shifts between post- and pre-replicative states during the cell cycle. We wanted to determine whether 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 LTV-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. Because the conversion of the origin footprinting profiles and origin firing, as assessed by two-dimensional 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.

Original languageEnglish
Pages (from-to)15881-15889
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number18
DOIs
Publication statusPublished - May 3 2002
Externally publishedYes

Fingerprint

Replication Origin
Yeast
Saccharomyces cerevisiae
Cell Cycle
Cells
S Phase
Origin Recognition Complex
Electrophoresis, Gel, Two-Dimensional
G1 Phase
Chromosomes
Electrophoresis
DNA Replication
Cell Division
Appointments and Schedules
Gels
Chemical activation
Association reactions
DNA

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Scheduled conversion of replication complex architecture at replication origins of Saccharomyces cerevisiae during the cell cycle. / Tadokoro, Ryusuke; Fujita, Masako; Miura, Hitoshi; Shirahige, Katsuhiko; Yoshikawa, Hiroshi; Tsurimoto, Toshiki; Obuse, Chikashi.

In: Journal of Biological Chemistry, Vol. 277, No. 18, 03.05.2002, p. 15881-15889.

Research output: Contribution to journalArticle

Tadokoro, Ryusuke ; Fujita, Masako ; Miura, Hitoshi ; Shirahige, Katsuhiko ; Yoshikawa, Hiroshi ; Tsurimoto, Toshiki ; Obuse, Chikashi. / Scheduled conversion of replication complex architecture at replication origins of Saccharomyces cerevisiae during the cell cycle. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 18. pp. 15881-15889.
@article{1d84d33d834348589e613add5d9490d5,
title = "Scheduled conversion of replication complex architecture at replication origins of Saccharomyces cerevisiae during the cell cycle",
abstract = "Replication of DNA within Saccharomyces 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 an origin recognition complex is formed at each origin and shifts between post- and pre-replicative states during the cell cycle. We wanted to determine whether 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 LTV-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. Because the conversion of the origin footprinting profiles and origin firing, as assessed by two-dimensional 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.",
author = "Ryusuke Tadokoro and Masako Fujita and Hitoshi Miura and Katsuhiko Shirahige and Hiroshi Yoshikawa and Toshiki Tsurimoto and Chikashi Obuse",
year = "2002",
month = "5",
day = "3",
doi = "10.1074/jbc.M200322200",
language = "English",
volume = "277",
pages = "15881--15889",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "18",

}

TY - JOUR

T1 - Scheduled conversion of replication complex architecture at replication origins of Saccharomyces cerevisiae during the cell cycle

AU - Tadokoro, Ryusuke

AU - Fujita, Masako

AU - Miura, Hitoshi

AU - Shirahige, Katsuhiko

AU - Yoshikawa, Hiroshi

AU - Tsurimoto, Toshiki

AU - Obuse, Chikashi

PY - 2002/5/3

Y1 - 2002/5/3

N2 - Replication of DNA within Saccharomyces 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 an origin recognition complex is formed at each origin and shifts between post- and pre-replicative states during the cell cycle. We wanted to determine whether 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 LTV-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. Because the conversion of the origin footprinting profiles and origin firing, as assessed by two-dimensional 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.

AB - Replication of DNA within Saccharomyces 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 an origin recognition complex is formed at each origin and shifts between post- and pre-replicative states during the cell cycle. We wanted to determine whether 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 LTV-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. Because the conversion of the origin footprinting profiles and origin firing, as assessed by two-dimensional 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.

UR - http://www.scopus.com/inward/record.url?scp=0037013261&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037013261&partnerID=8YFLogxK

U2 - 10.1074/jbc.M200322200

DO - 10.1074/jbc.M200322200

M3 - Article

VL - 277

SP - 15881

EP - 15889

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 18

ER -