Cooperative Cluster Formation, DNA Bending and Base-Flipping by O6-Alkylguanine-DNA Alkyltransferase
Abstract
O6-Alkylguanine-DNA alkyltransferase (AGT) repairs mutagenic O6-alkylguanine and O4-alkylthymine adducts in DNA, protecting the genome and also contributing to the resistance of tumors to chemotherapeutic alkylating agents. AGT binds DNA cooperatively, and cooperative interactions are likely to be important in lesion search and repair. We examined morphologies of complexes on long, unmodified DNAs, using analytical ultracentrifugation and atomic force microscopy. AGT formed clusters of ≤11 proteins. Longer clusters, predicted by the McGhee–von Hippel model, were not seen even at high [protein]. Interestingly, torsional stress due to DNA unwinding has the potential to limit cluster size to the observed range. DNA at cluster sites showed bend angles (∼0, ∼30 and ∼60°) that are consistent with models in which each protein induces a bend of ∼30°. Distributions of complexes along the DNA are incompatible with sequence specificity but suggest modest preference for DNA ends. These properties tell us about environments in which AGT may function. Small cooperative clusters and the ability to accommodate a range of DNA bends allow function where DNA topology is constrained, such as near DNA-replication complexes. The low sequence specificity allows efficient and unbiased lesion search across the entire genome.
Document Type
Article
Publication Date
2012
Digital Object Identifier (DOI)
http://dx.doi.org/10.1093/nar/gks574
Repository Citation
Tessmer, Ingrid; Melikishvili, Manana; and Fried, Michael G., "Cooperative Cluster Formation, DNA Bending and Base-Flipping by O6-Alkylguanine-DNA Alkyltransferase" (2012). Center for Structural Biology Faculty Publications. 7.
https://uknowledge.uky.edu/csb_facpub/7
Supplementary Data
Notes/Citation Information
Published in Nucleic Acids Research, v. 40, no. 17, p. 8296-8308.
© The Author(s) 2012. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.