Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB
The AlkB enzyme is an Fe(II)- and α-ketoglutarate-dependent dioxygenase that repairs DNA alkyl lesions by a direct reversal of damage mechanism as part of the adaptive response in E. coli. The reported substrate scope of AlkB includes simple DNA alkyl adducts, such as 1-methyladenine, 3-methylcytosi...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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American Chemical Society
2013
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| Online Access: | http://hdl.handle.net/1721.1/82033 https://orcid.org/0000-0001-5486-2755 https://orcid.org/0000-0001-6159-0778 https://orcid.org/0000-0002-2196-5691 |
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| author | Li, Deyu Fedeles, Bogdan I. Shrivastav, Nidhi Delaney, James C. Yang, Xuedong Wong, Cintyu Drennan, Catherine L. Essigmann, John M. |
| author2 | Massachusetts Institute of Technology. Center for Environmental Health Sciences |
| author_facet | Massachusetts Institute of Technology. Center for Environmental Health Sciences Li, Deyu Fedeles, Bogdan I. Shrivastav, Nidhi Delaney, James C. Yang, Xuedong Wong, Cintyu Drennan, Catherine L. Essigmann, John M. |
| author_sort | Li, Deyu |
| collection | MIT |
| description | The AlkB enzyme is an Fe(II)- and α-ketoglutarate-dependent dioxygenase that repairs DNA alkyl lesions by a direct reversal of damage mechanism as part of the adaptive response in E. coli. The reported substrate scope of AlkB includes simple DNA alkyl adducts, such as 1-methyladenine, 3-methylcytosine, 3-ethylcytosine, 1-methylguanine, 3-methylthymine, and N6-methyladenine, as well as more complex DNA adducts, such as 1,N6-ethenoadenine, 3,N4-ethenocytosine, and 1,N6-ethanoadenine. Previous studies have revealed, in a piecemeal way, that AlkB has an impressive repertoire of substrates. The present study makes two additions to this list, showing that alkyl adducts on the N2 position of guanine and N4 position of cytosine are also substrates for AlkB. Using high resolution ESI-TOF mass spectrometry, we show that AlkB has the biochemical capability to repair in vitro N2-methylguanine, N2-ethylguanine, N2-furan-2-yl-methylguanine, N2-tetrahydrofuran-2-yl-methylguanine, and N4-methylcytosine in ssDNA but not in dsDNA. When viewed together with previous work, the experimental data herein demonstrate that AlkB is able to repair all simple N-alkyl adducts occurring at the Watson–Crick base pairing interface of the four DNA bases, confirming AlkB as a versatile gatekeeper of genomic integrity under alkylation stress. |
| first_indexed | 2024-09-23T10:38:11Z |
| format | Article |
| id | mit-1721.1/82033 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:38:11Z |
| publishDate | 2013 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/820332022-09-27T13:52:47Z Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB Removal of N-Alkyl Modifications from N2-Alkylguanine and N4-Alkylcytosine in DNA by the Adaptive Response Protein AlkB Li, Deyu Fedeles, Bogdan I. Shrivastav, Nidhi Delaney, James C. Yang, Xuedong Wong, Cintyu Drennan, Catherine L. Essigmann, John M. Massachusetts Institute of Technology. Center for Environmental Health Sciences Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemistry Li, Deyu Fedeles, Bogdan I. Shrivastav, Nidhi Delaney, James C. Yang, Xuedong Wong, Cintyu Drennan, Catherine L. Essigmann, John M. The AlkB enzyme is an Fe(II)- and α-ketoglutarate-dependent dioxygenase that repairs DNA alkyl lesions by a direct reversal of damage mechanism as part of the adaptive response in E. coli. The reported substrate scope of AlkB includes simple DNA alkyl adducts, such as 1-methyladenine, 3-methylcytosine, 3-ethylcytosine, 1-methylguanine, 3-methylthymine, and N6-methyladenine, as well as more complex DNA adducts, such as 1,N6-ethenoadenine, 3,N4-ethenocytosine, and 1,N6-ethanoadenine. Previous studies have revealed, in a piecemeal way, that AlkB has an impressive repertoire of substrates. The present study makes two additions to this list, showing that alkyl adducts on the N2 position of guanine and N4 position of cytosine are also substrates for AlkB. Using high resolution ESI-TOF mass spectrometry, we show that AlkB has the biochemical capability to repair in vitro N2-methylguanine, N2-ethylguanine, N2-furan-2-yl-methylguanine, N2-tetrahydrofuran-2-yl-methylguanine, and N4-methylcytosine in ssDNA but not in dsDNA. When viewed together with previous work, the experimental data herein demonstrate that AlkB is able to repair all simple N-alkyl adducts occurring at the Watson–Crick base pairing interface of the four DNA bases, confirming AlkB as a versatile gatekeeper of genomic integrity under alkylation stress. National Institutes of Health (U.S.) (NIH grant ES002109) National Institutes of Health (U.S.) (NIH grant CA080024) National Institutes of Health (U.S.) (NIH grant CA26731) Howard Hughes Medical Institute (Investigator) 2013-11-07T21:07:10Z 2013-11-07T21:07:10Z 2013-08 2013-03 Article http://purl.org/eprint/type/JournalArticle 0893-228X 1520-5010 http://hdl.handle.net/1721.1/82033 Li, Deyu, Bogdan I. Fedeles, Nidhi Shrivastav, James C. Delaney, Xuedong Yang, Cintyu Wong, Catherine L. Drennan, and John M. Essigmann. “Removal of N-Alkyl Modifications from N2-Alkylguanine and N4-Alkylcytosine in DNA by the Adaptive Response Protein AlkB.” Chemical Research in Toxicology 26, no. 8 (August 19, 2013): 1182-1187. https://orcid.org/0000-0001-5486-2755 https://orcid.org/0000-0001-6159-0778 https://orcid.org/0000-0002-2196-5691 en_US http://dx.doi.org/10.1021/tx400096m Chemical Research in Toxicology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society ACS Author Choice |
| spellingShingle | Li, Deyu Fedeles, Bogdan I. Shrivastav, Nidhi Delaney, James C. Yang, Xuedong Wong, Cintyu Drennan, Catherine L. Essigmann, John M. Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB |
| title | Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB |
| title_full | Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB |
| title_fullStr | Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB |
| title_full_unstemmed | Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB |
| title_short | Removal of N-Alkyl Modifications from N[superscript 2]-Alkylguanine and N[superscript 4]-Alkylcytosine in DNA by the Adaptive Response Protein AlkB |
| title_sort | removal of n alkyl modifications from n superscript 2 alkylguanine and n superscript 4 alkylcytosine in dna by the adaptive response protein alkb |
| url | http://hdl.handle.net/1721.1/82033 https://orcid.org/0000-0001-5486-2755 https://orcid.org/0000-0001-6159-0778 https://orcid.org/0000-0002-2196-5691 |
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