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In vitro replication and repair of DNA containing a C2'-oxidized abasic site.
Biochemistry. 2004 Dec 07; 43(48):15217-22.B

Abstract

Abasic lesions are unable to form Watson-Crick hydrogen bonds with nucleotides. Nonetheless, polymerase and repair enzymes distinguish between various oxidized abasic lesions, as well as from nonoxidized abasic sites (AP). The C2-AP lesion is produced when DNA is exposed to gamma-radiolysis. Its effects on polymerases and repair enzymes are unknown. A recently reported method for the chemical synthesis of oligonucleotides containing C2-AP at a defined site was utilized for studying the activity of Klenow exo(-) and repair enzymes on templates containing the lesion. The C2-AP lesion has a similar effect on Klenow exo(-) as do AP and C4-AP sites. Deoxyadenosine is preferentially incorporated opposite C2-AP, but extension of the primer past the lesion is strongly blocked. C2-AP is incised less efficiently by exonuclease III and endonuclease IV than are other abasic lesions. Furthermore, although a Schiff base between C2-AP and endonuclease III can be chemically trapped, the location of the 3'-phosphate alpha with respect to the aldehyde prevents beta-elimination associated with the lyase activity of type I base excision repair enzymes. The interactions of the C2'-oxidized abasic site with Klenow exo(-) and repair enzymes suggest that the lesion will be mutagenic and that it will be removed by strand displacement synthesis and flap endonuclease processing via a long patch repair mechanism.

Authors+Show Affiliations

Department of Chemistry, The Johns Hopkins University, 3400 North Charles Street, Baltimore, Maryland 21218, USA. mgreenberg@jhu.eduNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15568814

Citation

Greenberg, Marc M., et al. "In Vitro Replication and Repair of DNA Containing a C2'-oxidized Abasic Site." Biochemistry, vol. 43, no. 48, 2004, pp. 15217-22.
Greenberg MM, Weledji YN, Kroeger KM, et al. In vitro replication and repair of DNA containing a C2'-oxidized abasic site. Biochemistry. 2004;43(48):15217-22.
Greenberg, M. M., Weledji, Y. N., Kroeger, K. M., & Kim, J. (2004). In vitro replication and repair of DNA containing a C2'-oxidized abasic site. Biochemistry, 43(48), 15217-22.
Greenberg MM, et al. In Vitro Replication and Repair of DNA Containing a C2'-oxidized Abasic Site. Biochemistry. 2004 Dec 7;43(48):15217-22. PubMed PMID: 15568814.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - In vitro replication and repair of DNA containing a C2'-oxidized abasic site. AU - Greenberg,Marc M, AU - Weledji,Yvonne N, AU - Kroeger,Kelly M, AU - Kim,Jaeseung, PY - 2004/12/1/pubmed PY - 2005/1/22/medline PY - 2004/12/1/entrez SP - 15217 EP - 22 JF - Biochemistry JO - Biochemistry VL - 43 IS - 48 N2 - Abasic lesions are unable to form Watson-Crick hydrogen bonds with nucleotides. Nonetheless, polymerase and repair enzymes distinguish between various oxidized abasic lesions, as well as from nonoxidized abasic sites (AP). The C2-AP lesion is produced when DNA is exposed to gamma-radiolysis. Its effects on polymerases and repair enzymes are unknown. A recently reported method for the chemical synthesis of oligonucleotides containing C2-AP at a defined site was utilized for studying the activity of Klenow exo(-) and repair enzymes on templates containing the lesion. The C2-AP lesion has a similar effect on Klenow exo(-) as do AP and C4-AP sites. Deoxyadenosine is preferentially incorporated opposite C2-AP, but extension of the primer past the lesion is strongly blocked. C2-AP is incised less efficiently by exonuclease III and endonuclease IV than are other abasic lesions. Furthermore, although a Schiff base between C2-AP and endonuclease III can be chemically trapped, the location of the 3'-phosphate alpha with respect to the aldehyde prevents beta-elimination associated with the lyase activity of type I base excision repair enzymes. The interactions of the C2'-oxidized abasic site with Klenow exo(-) and repair enzymes suggest that the lesion will be mutagenic and that it will be removed by strand displacement synthesis and flap endonuclease processing via a long patch repair mechanism. SN - 0006-2960 UR - https://www.unboundmedicine.com/medline/citation/15568814/In_vitro_replication_and_repair_of_DNA_containing_a_C2'_oxidized_abasic_site_ L2 - https://dx.doi.org/10.1021/bi048360c DB - PRIME DP - Unbound Medicine ER -