Acidic phospholipids inhibit the DNA‐binding activity of DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli
Summary In order to initiate chromosomal DNA replication in Escherichia coli, the DnaA protein must bind to both ATP and the origin of replication (oriC). Acidic phospholipids are known to inhibit DnaA binding to ATP, and here we examine the effects of various phospholipids on DnaA binding to oriC....
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| Veröffentlicht in: | Molecular microbiology 2002-10, Vol.46 (1), p.245-256 |
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| creator | Makise, Masaki Mima, Shinji Katsu, Takashi Tsuchiya, Tomofusa Mizushima, Tohru |
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In order to initiate chromosomal DNA replication in Escherichia coli, the DnaA protein must bind to both ATP and the origin of replication (oriC). Acidic phospholipids are known to inhibit DnaA binding to ATP, and here we examine the effects of various phospholipids on DnaA binding to oriC. Among the phospholipids in E. coli membrane, cardiolipin showed the strongest inhibition of DnaA binding to oriC. Synthetic phosphatidylglycerol containing unsaturated fatty acids inhibited binding more potently than did synthetic phosphatidylglycerol containing saturated fatty acids, suggesting that membrane fluidity is important. Thus, acidic phospholipids seem to inhibit DnaA binding to both oriC and adenine nucleotides in the same manner. Adenine nucleotides bound to DnaA did not affect the inhibitory effect of cardiolipin on DnaA binding to oriC. A mobility‐shift assay re‐vealed that acidic phospholipids inhibited formation of a DnaA–oriC complex containing several DnaA molecules. DNase I footprinting of DnaA binding to oriC showed that two DnaA binding sites (R2 and R3) were more sensitive to cardiolipin than other DnaA binding sites. Based on these in vitro data, the physiological relevance of this inhibitory effect of acidic phospholipids on DnaA binding to oriC is discussed. |
| doi_str_mv | 10.1046/j.1365-2958.2002.03161.x |
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In order to initiate chromosomal DNA replication in Escherichia coli, the DnaA protein must bind to both ATP and the origin of replication (oriC). Acidic phospholipids are known to inhibit DnaA binding to ATP, and here we examine the effects of various phospholipids on DnaA binding to oriC. Among the phospholipids in E. coli membrane, cardiolipin showed the strongest inhibition of DnaA binding to oriC. Synthetic phosphatidylglycerol containing unsaturated fatty acids inhibited binding more potently than did synthetic phosphatidylglycerol containing saturated fatty acids, suggesting that membrane fluidity is important. Thus, acidic phospholipids seem to inhibit DnaA binding to both oriC and adenine nucleotides in the same manner. Adenine nucleotides bound to DnaA did not affect the inhibitory effect of cardiolipin on DnaA binding to oriC. A mobility‐shift assay re‐vealed that acidic phospholipids inhibited formation of a DnaA–oriC complex containing several DnaA molecules. DNase I footprinting of DnaA binding to oriC showed that two DnaA binding sites (R2 and R3) were more sensitive to cardiolipin than other DnaA binding sites. Based on these in vitro data, the physiological relevance of this inhibitory effect of acidic phospholipids on DnaA binding to oriC is discussed.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1046/j.1365-2958.2002.03161.x</identifier><identifier>PMID: 12366847</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Science, Ltd</publisher><subject>Bacterial Proteins - metabolism ; Binding Sites - drug effects ; Cell Membrane - chemistry ; DNA Footprinting ; DNA Replication ; DNA, Bacterial - metabolism ; DNA-Binding Proteins - metabolism ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Origin Recognition Complex ; Phospholipids - chemistry ; Phospholipids - pharmacology ; Viral Proteins - metabolism</subject><ispartof>Molecular microbiology, 2002-10, Vol.46 (1), p.245-256</ispartof><rights>Copyright Blackwell Scientific Publications Ltd. Oct 2002</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5391-183fc3878be72f9f6d2a1867b17beec878ee219087c5de25ed01dc2f40c0d8e93</citedby><cites>FETCH-LOGICAL-c5391-183fc3878be72f9f6d2a1867b17beec878ee219087c5de25ed01dc2f40c0d8e93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1046%2Fj.1365-2958.2002.03161.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1046%2Fj.1365-2958.2002.03161.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,1433,27924,27925,45574,45575,46409,46833</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12366847$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Makise, Masaki</creatorcontrib><creatorcontrib>Mima, Shinji</creatorcontrib><creatorcontrib>Katsu, Takashi</creatorcontrib><creatorcontrib>Tsuchiya, Tomofusa</creatorcontrib><creatorcontrib>Mizushima, Tohru</creatorcontrib><title>Acidic phospholipids inhibit the DNA‐binding activity of DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>Summary
In order to initiate chromosomal DNA replication in Escherichia coli, the DnaA protein must bind to both ATP and the origin of replication (oriC). Acidic phospholipids are known to inhibit DnaA binding to ATP, and here we examine the effects of various phospholipids on DnaA binding to oriC. Among the phospholipids in E. coli membrane, cardiolipin showed the strongest inhibition of DnaA binding to oriC. Synthetic phosphatidylglycerol containing unsaturated fatty acids inhibited binding more potently than did synthetic phosphatidylglycerol containing saturated fatty acids, suggesting that membrane fluidity is important. Thus, acidic phospholipids seem to inhibit DnaA binding to both oriC and adenine nucleotides in the same manner. Adenine nucleotides bound to DnaA did not affect the inhibitory effect of cardiolipin on DnaA binding to oriC. A mobility‐shift assay re‐vealed that acidic phospholipids inhibited formation of a DnaA–oriC complex containing several DnaA molecules. DNase I footprinting of DnaA binding to oriC showed that two DnaA binding sites (R2 and R3) were more sensitive to cardiolipin than other DnaA binding sites. Based on these in vitro data, the physiological relevance of this inhibitory effect of acidic phospholipids on DnaA binding to oriC is discussed.</description><subject>Bacterial Proteins - metabolism</subject><subject>Binding Sites - drug effects</subject><subject>Cell Membrane - chemistry</subject><subject>DNA Footprinting</subject><subject>DNA Replication</subject><subject>DNA, Bacterial - metabolism</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Origin Recognition Complex</subject><subject>Phospholipids - chemistry</subject><subject>Phospholipids - pharmacology</subject><subject>Viral Proteins - metabolism</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkctu1DAUhi1ERYfCKyCLBSsSfMnFWbAYtaVUamEDEjvLcU7IGSVxsDOls2PLrs_Ik-B0RkViAwvLls53_mP7I4RylnKWFW82KZdFnogqV6lgTKRM8oKnt4_I6qHwmKxYlbNEKvHlmDwNYcMYl6yQT8gxF7IoVFauyM-1xQYtnToX4upxwiZQHDuscaZzB_Tsw_rXj7saxwbHr9TYGW9w3lHX0rPRrOnk3Qw4vr5nccQZzez8Uradd4MLbjD9EkI9TD1aM6MbI0jPg-3Ao-3QUBsHPyNHrekDPD_sJ-Tzu_NPp--Tq48Xl6frq8TmsuIJV7K1UpWqhlK0VVs0wnBVlDUvawAbCwCCV0yVNm9A5NAw3ljRZsyyRkElT8irfW68-bcthFkPGCz0vRnBbYMuBc9LmbF_glzl8UNlFsGXf4Ebt_VjfITmVZGzLGMLpPaQ9S4ED62ePA7G7zRnepGqN3pxpxd3epGq76Xq29j64pC_rQdo_jQeLEbg7R74jj3s_jtYX19fLif5Gw5lsiA</recordid><startdate>200210</startdate><enddate>200210</enddate><creator>Makise, Masaki</creator><creator>Mima, Shinji</creator><creator>Katsu, Takashi</creator><creator>Tsuchiya, Tomofusa</creator><creator>Mizushima, Tohru</creator><general>Blackwell Science, Ltd</general><general>Blackwell Publishing 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>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>200210</creationdate><title>Acidic phospholipids inhibit the DNA‐binding activity of DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli</title><author>Makise, Masaki ; Mima, Shinji ; Katsu, Takashi ; Tsuchiya, Tomofusa ; Mizushima, Tohru</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5391-183fc3878be72f9f6d2a1867b17beec878ee219087c5de25ed01dc2f40c0d8e93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Bacterial Proteins - metabolism</topic><topic>Binding Sites - drug effects</topic><topic>Cell Membrane - chemistry</topic><topic>DNA Footprinting</topic><topic>DNA Replication</topic><topic>DNA, Bacterial - metabolism</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Origin Recognition Complex</topic><topic>Phospholipids - chemistry</topic><topic>Phospholipids - pharmacology</topic><topic>Viral Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Makise, Masaki</creatorcontrib><creatorcontrib>Mima, Shinji</creatorcontrib><creatorcontrib>Katsu, Takashi</creatorcontrib><creatorcontrib>Tsuchiya, Tomofusa</creatorcontrib><creatorcontrib>Mizushima, Tohru</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Makise, Masaki</au><au>Mima, Shinji</au><au>Katsu, Takashi</au><au>Tsuchiya, Tomofusa</au><au>Mizushima, Tohru</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Acidic phospholipids inhibit the DNA‐binding activity of DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>2002-10</date><risdate>2002</risdate><volume>46</volume><issue>1</issue><spage>245</spage><epage>256</epage><pages>245-256</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><abstract>Summary
In order to initiate chromosomal DNA replication in Escherichia coli, the DnaA protein must bind to both ATP and the origin of replication (oriC). Acidic phospholipids are known to inhibit DnaA binding to ATP, and here we examine the effects of various phospholipids on DnaA binding to oriC. Among the phospholipids in E. coli membrane, cardiolipin showed the strongest inhibition of DnaA binding to oriC. Synthetic phosphatidylglycerol containing unsaturated fatty acids inhibited binding more potently than did synthetic phosphatidylglycerol containing saturated fatty acids, suggesting that membrane fluidity is important. Thus, acidic phospholipids seem to inhibit DnaA binding to both oriC and adenine nucleotides in the same manner. Adenine nucleotides bound to DnaA did not affect the inhibitory effect of cardiolipin on DnaA binding to oriC. A mobility‐shift assay re‐vealed that acidic phospholipids inhibited formation of a DnaA–oriC complex containing several DnaA molecules. DNase I footprinting of DnaA binding to oriC showed that two DnaA binding sites (R2 and R3) were more sensitive to cardiolipin than other DnaA binding sites. Based on these in vitro data, the physiological relevance of this inhibitory effect of acidic phospholipids on DnaA binding to oriC is discussed.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science, Ltd</pub><pmid>12366847</pmid><doi>10.1046/j.1365-2958.2002.03161.x</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Bacterial Proteins - metabolism Binding Sites - drug effects Cell Membrane - chemistry DNA Footprinting DNA Replication DNA, Bacterial - metabolism DNA-Binding Proteins - metabolism Escherichia coli - genetics Escherichia coli - metabolism Origin Recognition Complex Phospholipids - chemistry Phospholipids - pharmacology Viral Proteins - metabolism |
| title | Acidic phospholipids inhibit the DNA‐binding activity of DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli |
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