T4 ENDONUCLEASE V: STRUCTURE-FUNCTION ANALYSES

Project: Research project

Project Details

Description

Specific interactions between proteins and primary DNA
sequences provide a basic framework through which numerous
cellular functions are mediated: control of gene expression,
initiation of DNA synthesis, genetic recombination, initiation of
DNA repair and restriction/modification and compaction of
nucleic acids. Within this framework of DNA-interactive
proteins, two major categories of proteins can be distinguished as
those which bind or modify DNA at very specific sequence arrays
and those which interact with necleic acids through a more
general mechanism in which they recognize common DNA
structural features or DNA abnormalities. In contrast to the highly sequence-dependent protein-DNA
associations (i.e. repressor molecules, polymerases), the DNA
repair enzymes comprise a group of proteins which do not interact
at specific arrays of DNA sequence but rather monitor DNA for
aberrations which may have been caused by thermal, radiation or
chemcial challenge or errors made during DNA replication. The
lack of precise sequence recognition of these enzymes for their
substrates does not minimize the requirement for precision in
locating and recognizing the DNA aberration. Rather it mandates
that the protein-DNA interactions occur through more
generalized mechanisms and yet maintain a high degree of
accuracy, even in the absence of invariant residue contacts. This
proposal centers on the elucidation of the relationship between
certain protein sequences which are found in one such DNA repair
enzyme, the T4 endonuclease V and the observed biological and
enzymatic functions of that protein. The determination of these
more generalized structure-function relationships may serve as a
working model from which extrapolations can be made to other
DNA reactive proteins. This goal will be accomplished through the following specific
aims: 1) an analysis of endonuclease V mutants which were
produced by either oligonucleotide site-directed mutagenesis or
region-specific mutagenesis of the cloned gene which codes for
endonuclease V; the effects of those alterations on enzymatic
functions both in vivo and in vitro will be determined, 2)
structural comparisons between endonuclease V and a functionally
related UV endonuclease, 3) footprinting studies to characterize
specific points of contact between the protein and DNA, and 4)
elucidation of the X-ray crystal structure for the T4 endonuclease
V.
StatusFinished
Effective start/end date6/1/878/31/12

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $292,477.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $319,990.00
  • National Institutes of Health: $284,308.00
  • National Institutes of Health: $346,233.00
  • National Institutes of Health: $292,837.00
  • National Institutes of Health
  • National Institutes of Health: $353,082.00
  • National Institutes of Health: $298,000.00
  • National Institutes of Health: $346,382.00
  • National Institutes of Health: $3,622.00
  • National Institutes of Health: $222,016.00
  • National Institutes of Health
  • National Institutes of Health: $375,435.00
  • National Institutes of Health
  • National Institutes of Health: $331,030.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

ASJC

  • Environmental Science(all)
  • Medicine(all)

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