title: Structure-function relationships governing activity and stability of a DNA alkylation damage repair thermostable protein.
abstract: Alkylated DNA-protein alkyltransferases repair alkylated DNA bases, which are among the most common DNA lesions, and are evolutionary conserved, from prokaryotes to higher eukaryotes. The human ortholog, hAGT, is involved in resistance to alkylating chemotherapy drugs. We report here on the alkylated DNA-protein alkyltransferase, SsOGT, from an archaeal species living at high temperature, a condition that enhances the harmful effect of DNA alkylation. The exceptionally high stability of SsOGT gave us the unique opportunity to perform structural and biochemical analysis of a protein of this class in its post-reaction form. This analysis, along with those performed on SsOGT in its ligand-free and DNA-bound forms, provides insights in the structure-function relationships of the protein before, during and after DNA repair, suggesting a molecular basis for DNA recognition, catalytic activity and protein post-reaction fate, and giving hints on the mechanism of alkylation-induced inactivation of this class of proteins.
authors.affiliation: Institute of Biosciences and Bioresources, National Research Council of Italy, Via P. Castellino 111, 80125 Naples, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Perugino
authors.affiliation: DiSF-Dipartimento di Scienze del Farmaco, University of Piemonte Orientale 'A. Avogadro', Via Bovio 6, 28100 Novara, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Miggiano
authors.affiliation: Institute of Biosciences and Bioresources, National Research Council of Italy, Via P. Castellino 111, 80125 Naples, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Serpe
authors.affiliation: Institute of Biosciences and Bioresources, National Research Council of Italy, Via P. Castellino 111, 80125 Naples, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Vettone
authors.affiliation: Institute of Biosciences and Bioresources, National Research Council of Italy, Via P. Castellino 111, 80125 Naples, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Valenti
authors.affiliation: DiSF-Dipartimento di Scienze del Farmaco, University of Piemonte Orientale 'A. Avogadro', Via Bovio 6, 28100 Novara, Italy.
authors.forename: Samarpita
authors.initials: S
authors.lastname: Lahiri
authors.affiliation: DiSF-Dipartimento di Scienze del Farmaco, University of Piemonte Orientale 'A. Avogadro', Via Bovio 6, 28100 Novara, Italy.
authors.forename: Franca
authors.initials: F
authors.lastname: Rossi
authors.affiliation: Institute of Biosciences and Bioresources, National Research Council of Italy, Via P. Castellino 111, 80125 Naples, Italy.
authors.forename: Mosè
authors.initials: M
authors.lastname: Rossi
authors.affiliation: DiSF-Dipartimento di Scienze del Farmaco, University of Piemonte Orientale 'A. Avogadro', Via Bovio 6, 28100 Novara, Italy.
authors.forename: Menico
authors.initials: M
authors.lastname: Rizzi
authors.affiliation: Institute of Biosciences and Bioresources, National Research Council of Italy, Via P. Castellino 111, 80125 Naples, Italy maria.ciaramella@ibbr.cnr.it.
authors.forename: Maria
authors.initials: M
authors.lastname: Ciaramella
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000478:Alkylation; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004260:DNA Repair; D004795:Enzyme Stability; D008958:Models, Molecular; D009154:Mutation; D013329:Structure-Activity Relationship; D048229:Sulfolobus solfataricus / Q000201:enzymology
chemical_list: D019843:Archaeal Proteins; D004247:DNA; D019883:Alkyl and Aryl Transferases; C050956:DNA alkyltransferase