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 language | English |
|---|---|
| Pages (from-to) | 15881-15889 |
| Number of pages | 9 |
| Journal | Journal of Biological Chemistry |
| Volume | 277 |
| Issue number | 18 |
| DOIs | |
| Publication status | Published - May 3 2002 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Cell Biology
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 journal › Article
}
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
C2 - 11842092
AN - SCOPUS:0037013261
VL - 277
SP - 15881
EP - 15889
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 18
ER -