title: Site-directed mutagenesis of the YCDTDS amino acid motif of the phi 29 DNA polymerase.
abstract: The Bacillus subtilis phage phi 29 DNA polymerase, involved in protein-primed viral DNA replication, contains amino acid consensus sequences common to other alpha-like DNA polymerases. Using site-directed mutagenesis we have studied the functional significance of the most conserved C-terminal segment mainly represented by the YCDTDS motif. A series of single point mutants has been constructed and the corresponding proteins have been overproduced and characterized. Measurements, on crude fractions, of the activity of the mutant proteins in the formation of the protein p3-dAMP initiation complex and in an in situ DNA polymerase assay, indicate that the YCDTDS domain is involved both in initiation and in elongation reactions.
authors.affiliation: Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Bernad
authors.forename: L
authors.initials: L
authors.lastname: Blanco
authors.forename: M
authors.initials: M
authors.lastname: Salas
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001435:Bacteriophages / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D004257:DNA Polymerase II / Q000235:genetics; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D014780:Viruses / Q000201:enzymology / Q000235:genetics
chemical_list: D004279:DNA, Viral; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase