The Bacillus subtilis Counterpart of the Mammalian 3-Methyladenine DNA Glycosylase Has Hypoxanthine and 1,N6-Ethenoadenine as Preferred Substrates
The AAG family of 3-methyladenine DNA glycosylases was initially thought to be limited to mammalian cells, but genome sequencing efforts have revealed the presence of homologous proteins in certain prokaryotic species as well. Here, we report the first molecular characterization of a functional prok...
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| Veröffentlicht in: | The Journal of biological chemistry 2004-04, Vol.279 (14), p.13601-13606 |
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| creator | Aamodt, Randi M Falnes, Pål Ø Johansen, Rune F Seeberg, Erling Bjørås, Magnar |
| description | The AAG family of 3-methyladenine DNA glycosylases was initially thought to be limited to mammalian cells, but genome sequencing
efforts have revealed the presence of homologous proteins in certain prokaryotic species as well. Here, we report the first
molecular characterization of a functional prokaryotic AAG homologue, i.e. YxlJ, termed bAag, from Bacillus subtilis . The B. subtilis aag gene was expressed in Escherichia coli , and the protein was purified to homogeneity. As expected, B. subtilis Aag was found to be a DNA glycosylase, which releases 3-alkylated purines and hypoxanthine, as well as the cyclic etheno
adduct 1, N 6 -ethenoadenine from DNA. However, kinetic analysis showed that bAag removed hypoxanthine much faster than human AAG with a
10-fold higher value for k cat , whereas the rate of excision of 1, N 6 -ethenoadenine was found to be similar. In contrast, it was found that bAag removes 3-methyladenine and 3-methylguanine â¼10â20
times more slowly than human AAG, and there was hardly any detectable excision of 7-methylguanine. It thus appears that bAag
has a minor role in the repair of DNA alkylation damage and an important role in preventing the mutagenic effects of deaminated
purines and cyclic etheno adducts in Bacillus subtilis . |
| doi_str_mv | 10.1074/jbc.M314277200 |
| format | Article |
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efforts have revealed the presence of homologous proteins in certain prokaryotic species as well. Here, we report the first
molecular characterization of a functional prokaryotic AAG homologue, i.e. YxlJ, termed bAag, from Bacillus subtilis . The B. subtilis aag gene was expressed in Escherichia coli , and the protein was purified to homogeneity. As expected, B. subtilis Aag was found to be a DNA glycosylase, which releases 3-alkylated purines and hypoxanthine, as well as the cyclic etheno
adduct 1, N 6 -ethenoadenine from DNA. However, kinetic analysis showed that bAag removed hypoxanthine much faster than human AAG with a
10-fold higher value for k cat , whereas the rate of excision of 1, N 6 -ethenoadenine was found to be similar. In contrast, it was found that bAag removes 3-methyladenine and 3-methylguanine â¼10â20
times more slowly than human AAG, and there was hardly any detectable excision of 7-methylguanine. It thus appears that bAag
has a minor role in the repair of DNA alkylation damage and an important role in preventing the mutagenic effects of deaminated
purines and cyclic etheno adducts in Bacillus subtilis .</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M314277200</identifier><identifier>PMID: 14729667</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>Adenine - analogs & derivatives ; Adenine - metabolism ; Alkylation ; Animals ; Bacillus subtilis - enzymology ; Bacillus subtilis - genetics ; Cloning, Molecular ; Deamination ; DNA Glycosylases - genetics ; DNA Glycosylases - metabolism ; DNA Methylation ; Escherichia coli - genetics ; Hypoxanthine - metabolism ; Mammals ; Molecular Sequence Data ; Mutation ; Open Reading Frames ; Sequence Homology, Amino Acid ; Substrate Specificity</subject><ispartof>The Journal of biological chemistry, 2004-04, Vol.279 (14), p.13601-13606</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c2070-959c4eaba67048a01b6eb28647d1f74a39bae7af09e4a6268aa09499e7521fa43</citedby><cites>FETCH-LOGICAL-c2070-959c4eaba67048a01b6eb28647d1f74a39bae7af09e4a6268aa09499e7521fa43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27922,27923</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14729667$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Aamodt, Randi M</creatorcontrib><creatorcontrib>Falnes, Pål Ø</creatorcontrib><creatorcontrib>Johansen, Rune F</creatorcontrib><creatorcontrib>Seeberg, Erling</creatorcontrib><creatorcontrib>Bjørås, Magnar</creatorcontrib><title>The Bacillus subtilis Counterpart of the Mammalian 3-Methyladenine DNA Glycosylase Has Hypoxanthine and 1,N6-Ethenoadenine as Preferred Substrates</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The AAG family of 3-methyladenine DNA glycosylases was initially thought to be limited to mammalian cells, but genome sequencing
efforts have revealed the presence of homologous proteins in certain prokaryotic species as well. Here, we report the first
molecular characterization of a functional prokaryotic AAG homologue, i.e. YxlJ, termed bAag, from Bacillus subtilis . The B. subtilis aag gene was expressed in Escherichia coli , and the protein was purified to homogeneity. As expected, B. subtilis Aag was found to be a DNA glycosylase, which releases 3-alkylated purines and hypoxanthine, as well as the cyclic etheno
adduct 1, N 6 -ethenoadenine from DNA. However, kinetic analysis showed that bAag removed hypoxanthine much faster than human AAG with a
10-fold higher value for k cat , whereas the rate of excision of 1, N 6 -ethenoadenine was found to be similar. In contrast, it was found that bAag removes 3-methyladenine and 3-methylguanine â¼10â20
times more slowly than human AAG, and there was hardly any detectable excision of 7-methylguanine. It thus appears that bAag
has a minor role in the repair of DNA alkylation damage and an important role in preventing the mutagenic effects of deaminated
purines and cyclic etheno adducts in Bacillus subtilis .</description><subject>Adenine - analogs & derivatives</subject><subject>Adenine - metabolism</subject><subject>Alkylation</subject><subject>Animals</subject><subject>Bacillus subtilis - enzymology</subject><subject>Bacillus subtilis - genetics</subject><subject>Cloning, Molecular</subject><subject>Deamination</subject><subject>DNA Glycosylases - genetics</subject><subject>DNA Glycosylases - metabolism</subject><subject>DNA Methylation</subject><subject>Escherichia coli - genetics</subject><subject>Hypoxanthine - metabolism</subject><subject>Mammals</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Open Reading Frames</subject><subject>Sequence Homology, Amino Acid</subject><subject>Substrate Specificity</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkE1P4zAQhi3ECgq7V47IB46k2I5rx0cohSK13ZWWlfYWjZMJMcpHZSeC_g1-MUYFMZeRRs_7avQQcsbZlDMtr55tMV2nXAqtBWMHZMJZlibpjP8_JBPGBE-MmGXH5CSEZxZHGn5EjrnUwiilJ-TtsUZ6A4VrmjHQMNrBNS7QeT92A_ot-IH2FR0itIa2hcZBR9NkjUO9a6DEznVIbzfX9L7ZFX2It4B0CYEud9v-Fbqh_gCgKym_3KhkEYu6_isXsT8eK_QeS_p3tGHwMGD4SX5U0AT89blPyb-7xeN8max-3z_Mr1dJIZhmiZmZQiJYUJrJDBi3Cq3IlNQlr7SE1FhADRUzKEEJlQEwI41BPRO8Apmekum-t_B9CPGRfOtdC36Xc5Z_yM2j3Pxbbgyc7wPb0bZYfuOfNiNwsQdq91S_OI-5dX1RY5sLbSKW81Qxnr4DjJyC2g</recordid><startdate>20040402</startdate><enddate>20040402</enddate><creator>Aamodt, Randi M</creator><creator>Falnes, Pål Ø</creator><creator>Johansen, Rune F</creator><creator>Seeberg, Erling</creator><creator>Bjørås, Magnar</creator><general>American Society for Biochemistry and Molecular Biology</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></search><sort><creationdate>20040402</creationdate><title>The Bacillus subtilis Counterpart of the Mammalian 3-Methyladenine DNA Glycosylase Has Hypoxanthine and 1,N6-Ethenoadenine as Preferred Substrates</title><author>Aamodt, Randi M ; Falnes, Pål Ø ; Johansen, Rune F ; Seeberg, Erling ; Bjørås, Magnar</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c2070-959c4eaba67048a01b6eb28647d1f74a39bae7af09e4a6268aa09499e7521fa43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Adenine - analogs & derivatives</topic><topic>Adenine - metabolism</topic><topic>Alkylation</topic><topic>Animals</topic><topic>Bacillus subtilis - enzymology</topic><topic>Bacillus subtilis - genetics</topic><topic>Cloning, Molecular</topic><topic>Deamination</topic><topic>DNA Glycosylases - genetics</topic><topic>DNA Glycosylases - metabolism</topic><topic>DNA Methylation</topic><topic>Escherichia coli - genetics</topic><topic>Hypoxanthine - metabolism</topic><topic>Mammals</topic><topic>Molecular Sequence Data</topic><topic>Mutation</topic><topic>Open Reading Frames</topic><topic>Sequence Homology, Amino Acid</topic><topic>Substrate Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Aamodt, Randi M</creatorcontrib><creatorcontrib>Falnes, Pål Ø</creatorcontrib><creatorcontrib>Johansen, Rune F</creatorcontrib><creatorcontrib>Seeberg, Erling</creatorcontrib><creatorcontrib>Bjørås, Magnar</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Aamodt, Randi M</au><au>Falnes, Pål Ø</au><au>Johansen, Rune F</au><au>Seeberg, Erling</au><au>Bjørås, Magnar</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Bacillus subtilis Counterpart of the Mammalian 3-Methyladenine DNA Glycosylase Has Hypoxanthine and 1,N6-Ethenoadenine as Preferred Substrates</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2004-04-02</date><risdate>2004</risdate><volume>279</volume><issue>14</issue><spage>13601</spage><epage>13606</epage><pages>13601-13606</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The AAG family of 3-methyladenine DNA glycosylases was initially thought to be limited to mammalian cells, but genome sequencing
efforts have revealed the presence of homologous proteins in certain prokaryotic species as well. Here, we report the first
molecular characterization of a functional prokaryotic AAG homologue, i.e. YxlJ, termed bAag, from Bacillus subtilis . The B. subtilis aag gene was expressed in Escherichia coli , and the protein was purified to homogeneity. As expected, B. subtilis Aag was found to be a DNA glycosylase, which releases 3-alkylated purines and hypoxanthine, as well as the cyclic etheno
adduct 1, N 6 -ethenoadenine from DNA. However, kinetic analysis showed that bAag removed hypoxanthine much faster than human AAG with a
10-fold higher value for k cat , whereas the rate of excision of 1, N 6 -ethenoadenine was found to be similar. In contrast, it was found that bAag removes 3-methyladenine and 3-methylguanine â¼10â20
times more slowly than human AAG, and there was hardly any detectable excision of 7-methylguanine. It thus appears that bAag
has a minor role in the repair of DNA alkylation damage and an important role in preventing the mutagenic effects of deaminated
purines and cyclic etheno adducts in Bacillus subtilis .</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>14729667</pmid><doi>10.1074/jbc.M314277200</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Adenine - analogs & derivatives Adenine - metabolism Alkylation Animals Bacillus subtilis - enzymology Bacillus subtilis - genetics Cloning, Molecular Deamination DNA Glycosylases - genetics DNA Glycosylases - metabolism DNA Methylation Escherichia coli - genetics Hypoxanthine - metabolism Mammals Molecular Sequence Data Mutation Open Reading Frames Sequence Homology, Amino Acid Substrate Specificity |
| title | The Bacillus subtilis Counterpart of the Mammalian 3-Methyladenine DNA Glycosylase Has Hypoxanthine and 1,N6-Ethenoadenine as Preferred Substrates |
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