MOLECULAR EFFECTORS OF ENZYMATIC DNA SYNTHESIS

Project Details

Description

Our long-range goals are to understand and unravel some of the intricacies
in the process of the enzymatic synthesis of DNA. Enzymes catalyzing DNA
synthesis require muliple components and exhibit several common and a few
unique properties. Yet the basis and/or components responsible for these
properties have not been identified. In this proposal, our intention is to
critically evaluate the process of substrate and template-primer binding
through the identification and structural analysis of the involved
functional domains using Klenow fragment of E. coli pol. I as a model
enzyme. A large body of information on the catalytic properties and
requirements for DNA synthesis, as well as the primary structure of this
enzyme are now known. The results obtained will not only clarify the basic
mechanisms of the DNA polymerase reaction but may provide insight into
properties which may have important practical applications. We have developed procedures for the covalent linkage of reagents to
specific enzyme sites and plan to develop new protocols and improve
existing protocols for the efficient labeling of binding sites. Our goal
is to isolate peptides bearing the substrate and template-primer binding
sites from Klenow fragment of E. coli Pol. I and to determine their primary
sequence. Thus, structural studies together with functional analyses of
various structural domains within individual DNA polymerases will clarify
the mechanisms of the complex multicomponent process of DNA synthesis.
StatusFinished
Effective start/end date8/5/858/31/06

Funding

  • National Institutes of Health: $274,750.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $274,750.00
  • National Institutes of Health: $274,750.00
  • National Institutes of Health
  • National Institutes of Health: $274,750.00

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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