The abasic site as a challenge to DNA polymerase
An abasic site in DNA creates a strong block to DNA polymerase and is a mutagenic base lesion. In this study, we present structural and dynamic properties of duplex oligodeoxynucleotides containing G, C and T opposite a model abasic site studied by one and two-dimensional nuclear magnetic resonance...
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| Published in | Journal of molecular biology Vol. 213; no. 2; pp. 303 - 314 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Elsevier Ltd
01.05.1990
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| Online Access | Get full text |
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| Abstract | An abasic site in DNA creates a strong block to DNA polymerase and is a mutagenic base lesion. In this study, we present structural and dynamic properties of duplex oligodeoxynucleotides containing G, C and T opposite a model abasic site studied by one and two-dimensional nuclear magnetic resonance spectroscopy. We have demonstrated that A opposite the abasic site was positioned within the helix as if paired with T, and that the A residue melted co-operatively with the surrounding helix. We report here that G opposite the abasic site is also observed to be predominantly intrahelical in a normal
anti conformation at low temperature. With increasing temperature, the mobility of the G residue increases rapidly and apparently is in a “melted state” well before denaturation of the helix. At low temperature, two species are found for T opposite the abasic site; one, intrahelical, one extrahelical. These species are in slow exchange with one another on a proton nuclear magnetic resonance time-scale. The two species then move into fast exchange with increasing temperature and the proportion of the extra-helical form increases. When C is positioned opposite the abasic site, both the C residue and the abasic sugar are extrahelical, the helix collapses, and the adjacent G·C base-pairs stack over one another. On the basis of these observations, we propose a model that explains why the abasic site acts to block DNA replication. Further, we suggest an explanation for the observed polymerase preference for base selection at abasic sites. |
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| AbstractList | An abasic site in DNA creates a strong block to DNA polymerase and is a mutagenic base lesion. In this study, we present structural and dynamic properties of duplex oligodeoxynucleotides containing G, C and T opposite a model abasic site studied by one and two-dimensional nuclear magnetic resonance spectroscopy. We have demonstrated that A opposite the abasic site was positioned within the helix as if paired with T, and that the A residue melted co-operatively with the surrounding helix. We report here that G opposite the abasic site is also observed to be predominantly intrahelical in a normal
anti conformation at low temperature. With increasing temperature, the mobility of the G residue increases rapidly and apparently is in a “melted state” well before denaturation of the helix. At low temperature, two species are found for T opposite the abasic site; one, intrahelical, one extrahelical. These species are in slow exchange with one another on a proton nuclear magnetic resonance time-scale. The two species then move into fast exchange with increasing temperature and the proportion of the extra-helical form increases. When C is positioned opposite the abasic site, both the C residue and the abasic sugar are extrahelical, the helix collapses, and the adjacent G·C base-pairs stack over one another. On the basis of these observations, we propose a model that explains why the abasic site acts to block DNA replication. Further, we suggest an explanation for the observed polymerase preference for base selection at abasic sites. |
| Author | Fazakerley, G.V. Guschlbauer, W. Kaplan, B.E. Cuniasse, Ph Sowers, L.C. |
| Author_xml | – sequence: 1 givenname: Ph surname: Cuniasse fullname: Cuniasse, Ph – sequence: 2 givenname: G.V. surname: Fazakerley fullname: Fazakerley, G.V. – sequence: 3 givenname: W. surname: Guschlbauer fullname: Guschlbauer, W. – sequence: 4 givenname: B.E. surname: Kaplan fullname: Kaplan, B.E. – sequence: 5 givenname: L.C. surname: Sowers fullname: Sowers, L.C. |
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| Snippet | An abasic site in DNA creates a strong block to DNA polymerase and is a mutagenic base lesion. In this study, we present structural and dynamic properties of... |
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| Title | The abasic site as a challenge to DNA polymerase |
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