title: Structural Basis for the KlenTaq DNA Polymerase Catalysed Incorporation of Alkene- versus Alkyne-Modified Nucleotides.
abstract: Efficient incorporation of modified nucleotides by DNA polymerases is essential for many cutting-edge biomolecular technologies. The present study compares the acceptance of either alkene- or alkyne-modified nucleotides by KlenTaq DNA polymerase and provides structural insights into how 7-deaza-adenosine and deoxyuridine with attached alkene-modifications are incorporated into the growing DNA strand. Thereby, we identified modified nucleotides that prove to be superior substrates for KlenTaq DNA polymerase compared with their natural analogues. The knowledge can be used to guide future design of functionalized nucleotide building blocks.
authors.affiliation: Department of Chemistry and Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Audrey
authors.initials: A
authors.lastname: Hottin
authors.affiliation: Department of Chemistry and Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Betz
authors.affiliation: Department of Chemistry and Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Kay
authors.initials: K
authors.lastname: Diederichs
authors.affiliation: Department of Chemistry and Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
mesh_terms: D000475:Alkenes / Q000737:chemistry*; D000480:Alkynes / Q000737:chemistry*; D055162:Biocatalysis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D003857:Deoxyuridine / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D021141:Nucleic Acid Amplification Techniques; D009711:Nucleotides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D014372:Tubercidin / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism
chemical_list: D000475:Alkenes; D000480:Alkynes; D009711:Nucleotides; D004259:DNA-Directed DNA Polymerase; D014372:Tubercidin; D003857:Deoxyuridine
keywords: DNA replication; DNA structures; labelling DNA polymerase; nucleotides; structure elucidation