Topologies of Complexes Containing O6-Alkylguanine-DNA Alkyltransferase and DNA

Claire A. Adams, Manana Melikishvili, David W. Rodgers, Joseph J. Rasimas, Anthony Pegg, Michael G. Fried

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The mutagenic and cytotoxic effects of many alkylating agents are reduced by O6-alkylguanine-DNA alkyltransferase (AGT). In humans, this protein not only protects the integrity of the genome, but also contributes to the resistance of tumors to DNA-alkylating chemotherapeutic agents. Here we describe and test models for cooperative multiprotein complexes of AGT with single-stranded and duplex DNAs that are based on in vitro binding data and the crystal structure of a 1:1 AGT-DNA complex. These models predict that cooperative assemblies contain a three-start helical array of proteins with dominant protein-protein interactions between the amino-terminal face of protein n and the carboxy-terminal face of protein n + 3, and they predict that binding duplex DNA does not require large changes in B-form DNA geometry. Experimental tests using protein cross-linking analyzed by mass spectrometry, electrophoretic and analytical ultracentrifugation binding assays, and topological analyses with closed circular DNA show that the properties of multiprotein AGT-DNA complexes are consistent with these predictions.

Original languageEnglish (US)
Pages (from-to)248-263
Number of pages16
JournalJournal of Molecular Biology
Volume389
Issue number2
DOIs
StatePublished - Jun 5 2009

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DNA
Proteins
Alkylating Agents
B-Form DNA
Circular DNA
Multiprotein Complexes
Protein Array Analysis
Single-Stranded DNA
Ultracentrifugation
DNA alkyltransferase
Mass Spectrometry
Genome
Neoplasms

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

Adams, C. A., Melikishvili, M., Rodgers, D. W., Rasimas, J. J., Pegg, A., & Fried, M. G. (2009). Topologies of Complexes Containing O6-Alkylguanine-DNA Alkyltransferase and DNA. Journal of Molecular Biology, 389(2), 248-263. https://doi.org/10.1016/j.jmb.2009.03.067
Adams, Claire A. ; Melikishvili, Manana ; Rodgers, David W. ; Rasimas, Joseph J. ; Pegg, Anthony ; Fried, Michael G. / Topologies of Complexes Containing O6-Alkylguanine-DNA Alkyltransferase and DNA. In: Journal of Molecular Biology. 2009 ; Vol. 389, No. 2. pp. 248-263.
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Adams, CA, Melikishvili, M, Rodgers, DW, Rasimas, JJ, Pegg, A & Fried, MG 2009, 'Topologies of Complexes Containing O6-Alkylguanine-DNA Alkyltransferase and DNA', Journal of Molecular Biology, vol. 389, no. 2, pp. 248-263. https://doi.org/10.1016/j.jmb.2009.03.067

Topologies of Complexes Containing O6-Alkylguanine-DNA Alkyltransferase and DNA. / Adams, Claire A.; Melikishvili, Manana; Rodgers, David W.; Rasimas, Joseph J.; Pegg, Anthony; Fried, Michael G.

In: Journal of Molecular Biology, Vol. 389, No. 2, 05.06.2009, p. 248-263.

Research output: Contribution to journalArticle

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