Project Details

Description

The recent discovery of reverse transcriptase (RT) in the prokaryotes opens
an exciting new biological system to study the origin of RT, and its roles
and functions in the prokaryotes. RT in myxobacteria appears to be as old
as other genomic genes, indicating that this enzyme existed before the
eukaryotes evolved. On the other hand, our data suggest that RTs in
Escherichia coli were acquired during the course of their evolution. In
both cases, RT is associated with a genetic system (retron), which
functions to produce a peculiar branched RNA-linked msDNA. In this proposal, we will use E. coli as a model system to elucidate their
functions and roles in the cells. For this purpose: (1) We will first
focus our efforts on further biochemical characterization of extensively
heterogeneous retrons from independent clinical E. coli isolates. We will
determine the entire genetic structure of these retrons including the DNA
sequence and how they are integrated in the E. coli genome. (2) In order
to elucidate the roles and functions of the retron, we will attempt to
determine how the msDNA molecules exist in the cells. For this purpose,
the protein-msDNA complex will be isolated and biochemically characterized,
and its possible role in the production of cDNA in the cells and its
ability to function as a transposable element will be examined. (3) We
will also attempt to purify RTs and other components required for the
synthesis of msDNA in order to elucidate the precise molecular events
leading to the production of msDNA molecules. (4) In order to elucidate
how many more natural E. coli isolates for the existence of retrons and to
establish their phylogenetic relationship. We will also examine the
existence of retrons or the production of msDNA in other prokaryotes, in
particular archaebacteria. (5) We will attempt to create artificial
retrons in E. coli which may be used for cDNA synthesis in vivo, for gene
amplification, for the production of stable ribozymes or as an easy
screening method to detect drugs against RT such as AZT.
StatusFinished
Effective start/end date4/1/903/31/01

Funding

  • National Institutes of Health: $122,774.00
  • National Institutes of Health
  • National Institutes of Health: $258,222.00
  • National Institutes of Health: $240,717.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $257,666.00
  • National Institutes of Health
  • National Institutes of Health: $268,334.00
  • National Institutes of Health

ASJC

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

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