Competition between the replication initiator DnaA and the sequestration factor SeqA for binding to the hemimethylated chromosomal origin of E. coli in vitro
Following replication initiation, the replication origin (oriC) in Escherichia coli enters a hemimethylated state at Dam methylation sites which are recognized by the SeqA protein. SeqA binds preferentially to hemimethylated GATC sequences of DNA in vitro. SeqA is essential for the synchronous initi...
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| Veröffentlicht in: | Genes to cells : devoted to molecular & cellular mechanisms 2000-11, Vol.5 (11), p.873-884 |
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| creator | Taghbalout, Aziz Landoulsi, Ahmed Kern, Renée Yamazoe, Mitsuyoshi Hiraga, Sota Holland, Barry Kohiyama, Masamichi Malki, Abderrahim |
| description | Following replication initiation, the replication origin (oriC) in Escherichia coli enters a hemimethylated state at Dam methylation sites which are recognized by the SeqA protein. SeqA binds preferentially to hemimethylated GATC sequences of DNA in vitro. SeqA is essential for the synchronous initiation of chromosome replication from oriC copies in vivo.
We show that: (i) purified SeqA binds AT‐rich and 13‐mers regions and two DnaA boxes, R1 and M, of hemimethylated oriC. (ii) SeqA inhibits the in vitro replication of a hemimethylated oriC plasmid more efficiently than the fully methylated, (iii) SeqA inhibits competitive binding of DnaA protein to the regions of the hemimethylated oriC plasmid, explaining the mechanism of its inhibitory effect. The inhibition of DnaA binding by SeqA also occurs efficiently on a small hemimethylated oriC fragment containing both R1 and M DnaA boxes, but not the 13‐mer region.
SeqA binds strongly the long region from the AT‐rich region to the M DnaA box of the hemimethylated oriC DNA and releases DnaA molecules from the long region. |
| doi_str_mv | 10.1046/j.1365-2443.2000.00380.x |
| format | Article |
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We show that: (i) purified SeqA binds AT‐rich and 13‐mers regions and two DnaA boxes, R1 and M, of hemimethylated oriC. (ii) SeqA inhibits the in vitro replication of a hemimethylated oriC plasmid more efficiently than the fully methylated, (iii) SeqA inhibits competitive binding of DnaA protein to the regions of the hemimethylated oriC plasmid, explaining the mechanism of its inhibitory effect. The inhibition of DnaA binding by SeqA also occurs efficiently on a small hemimethylated oriC fragment containing both R1 and M DnaA boxes, but not the 13‐mer region.
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We show that: (i) purified SeqA binds AT‐rich and 13‐mers regions and two DnaA boxes, R1 and M, of hemimethylated oriC. (ii) SeqA inhibits the in vitro replication of a hemimethylated oriC plasmid more efficiently than the fully methylated, (iii) SeqA inhibits competitive binding of DnaA protein to the regions of the hemimethylated oriC plasmid, explaining the mechanism of its inhibitory effect. The inhibition of DnaA binding by SeqA also occurs efficiently on a small hemimethylated oriC fragment containing both R1 and M DnaA boxes, but not the 13‐mer region.
SeqA binds strongly the long region from the AT‐rich region to the M DnaA box of the hemimethylated oriC DNA and releases DnaA molecules from the long region.</description><subject>AT Rich Sequence - physiology</subject><subject>Bacterial Outer Membrane Proteins</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacterial Proteins - pharmacology</subject><subject>Binding Sites - genetics</subject><subject>Binding, Competitive - genetics</subject><subject>DNA Methylation - drug effects</subject><subject>DNA Replication - drug effects</subject><subject>DNA Replication - genetics</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>DnaA protein</subject><subject>Escherichia coli</subject><subject>Escherichia coli Proteins</subject><subject>Origin Recognition Complex</subject><subject>Plasmids - genetics</subject><subject>Promoter Regions, Genetic - genetics</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Regulatory Sequences, Nucleic Acid - genetics</subject><subject>Replication Origin - genetics</subject><subject>replication origins</subject><subject>SeqA protein</subject><subject>Site-Specific DNA-Methyltransferase (Adenine-Specific) - antagonists & inhibitors</subject><subject>Transcription Factors</subject><subject>Viral Proteins - genetics</subject><issn>1356-9597</issn><issn>1365-2443</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1v1DAQhiMEoqXlLyCfuCX4Mx8Sl9VSClKlHmjPlmOPu14l8db20u6P6X_Fya7gSE9-5XlmRqOnKBDBFcG8_rKtCKtFSTlnFcUYVxizFlfPb4rzv4W3cxZ12YmuOSs-xLjFmDCKxfvijBBCKWvEefGy9uMOkkvOT6iH9AQwobQBFGA3OK2Wfzfluko-oG-TWiE1mQWJ8LiHmMIRskrPxC94XCGbQ-8m46YHlPwCb2B0I6TNYVAJDNKb4Ecf_agG5IN7cBPyFl1VSPvB5YXot0vBXxbvrBoifDy9F8X996u79Y_y5vb653p1U2rOBS4ZNgSbjqmGCqBcK2Pbuqaqxb3R1NYN5UQbYwnRttZ93ynR6V51XOnOip6zi-Lzce4u-OUmObqoYRjUBH4fZZ7LCCf4vyBp2qyiIxlsj6AOPsYAVu6CG1U4SILl7FBu5axKzqrk7FAuDuVzbv102rHvRzD_Gk_SMvD1CDy5AQ6vHiyv79Y5sD84va3d</recordid><startdate>200011</startdate><enddate>200011</enddate><creator>Taghbalout, Aziz</creator><creator>Landoulsi, Ahmed</creator><creator>Kern, Renée</creator><creator>Yamazoe, Mitsuyoshi</creator><creator>Hiraga, Sota</creator><creator>Holland, Barry</creator><creator>Kohiyama, Masamichi</creator><creator>Malki, Abderrahim</creator><general>Blackwell Science Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>200011</creationdate><title>Competition between the replication initiator DnaA and the sequestration factor SeqA for binding to the hemimethylated chromosomal origin of E. coli in vitro</title><author>Taghbalout, Aziz ; 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SeqA binds preferentially to hemimethylated GATC sequences of DNA in vitro. SeqA is essential for the synchronous initiation of chromosome replication from oriC copies in vivo.
We show that: (i) purified SeqA binds AT‐rich and 13‐mers regions and two DnaA boxes, R1 and M, of hemimethylated oriC. (ii) SeqA inhibits the in vitro replication of a hemimethylated oriC plasmid more efficiently than the fully methylated, (iii) SeqA inhibits competitive binding of DnaA protein to the regions of the hemimethylated oriC plasmid, explaining the mechanism of its inhibitory effect. The inhibition of DnaA binding by SeqA also occurs efficiently on a small hemimethylated oriC fragment containing both R1 and M DnaA boxes, but not the 13‐mer region.
SeqA binds strongly the long region from the AT‐rich region to the M DnaA box of the hemimethylated oriC DNA and releases DnaA molecules from the long region.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>11122375</pmid><doi>10.1046/j.1365-2443.2000.00380.x</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Genes to cells : devoted to molecular & cellular mechanisms, 2000-11, Vol.5 (11), p.873-884 |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Wiley Free Content; Open Access Titles of Japan |
| subjects | AT Rich Sequence - physiology Bacterial Outer Membrane Proteins Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacterial Proteins - pharmacology Binding Sites - genetics Binding, Competitive - genetics DNA Methylation - drug effects DNA Replication - drug effects DNA Replication - genetics DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism DnaA protein Escherichia coli Escherichia coli Proteins Origin Recognition Complex Plasmids - genetics Promoter Regions, Genetic - genetics Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Regulatory Sequences, Nucleic Acid - genetics Replication Origin - genetics replication origins SeqA protein Site-Specific DNA-Methyltransferase (Adenine-Specific) - antagonists & inhibitors Transcription Factors Viral Proteins - genetics |
| title | Competition between the replication initiator DnaA and the sequestration factor SeqA for binding to the hemimethylated chromosomal origin of E. coli in vitro |
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