Use of genetic analyses to probe structure, function, and dynamics of bacteriophage T4 DNA polymerase.

Functionally distinct mutant DNA polymerases have been isolated by the genetic selection strategies described here. These methods can be supplemented by the use of targeted mutagenesis procedures to enhance mutagenesis of DNA polymerase genes and to direct mutagenesis to specific sites in cloned DNA polymerases (see [22-24, 28], this volume). The power of genetic selection is in the ability to identify amino acid residues that are critical for protein structure and function that may not be obvious from studies of structural data alone. For the study of DNA polymerases, it is essential to identify residues involved in the movement of the DNA polymerase along the DNA template and in shuttling the DNA between the polymerase and exonuclease active centers. Ongoing studies are directed toward these goals.