Project Details
Description
DESCRIPTION (adapted from applicant's abstract): The line of inquiry proposed
in this research program is based on the hypothesis that the mitochondrial
ATP-dependent Lon protease monitors both the integrity and expression of the
mitochondrial genome as well as the folding and assembly of nascent
polypeptides. The goals of this proposal are to analyze the structure and
function of yeast Lon and to identify mammalian and yeast proteins that
functionally overlap with Lon. A combination of molecular biology, genetics,
biochemistry and structural biology will be used to address the following aims:
To analyze the role of Lon in regulating the mtDNA-protein interactions. The
DNA-binding function of Lon is conserved from bacteria to humans, however the
physiological importance of this activity is not understood. The investigator
will identify the endogenous mtDNA sequences bound by yeast Lon, analyze the
role of Lon in mtDNA-protein complexes called nucleoids and identify the
protein(s) responsible for increased double-stranded DNA-binding in
mitochondria that lack Lon.
To identify and characterize mammalian and yeast proteins that have a shared
function with Lon. Genetic complementation of a temperature-sensitive Lon
mutant using mammalian libraries designed for expression in yeast will be
carried out. The mammalian proteins identified will be characterized in
mammalian cell systems. In addition, a genome-wide survey of transcript level
changes in yeast deleted for the Lon gene will be conducted using
oligonucleotide probe microarrays.
To elucidate the structural dynamics and function of the homo-heptameric
ring-shaped Lon protease using circular dichroism and high-resolution electron
microscopy.
Mitochondrial dysfunction has been implicated in a wide variety of degenerative
disorders, aging and cancer. The best-characterized genetic mutations that lead
to a loss of mitochondrial function are deletions, point mutations and base
substitutions in mitochondrial DNA. Cellular responses to stress caused by
oxidative damage and aging are likely to involve the action of ATP-dependent
proteases which selectively degrade denatured, aggregated and unassembled
proteins. Understanding the molecular details of how ATP-dependent proteases
ensure cellular homeostasis by monitoring protein biogenesis as well as the
integrity and expression of the mitochondrial genome, will provide insight into
mechanisms that help to prevent mitochondrial dysfunction.
Status | Finished |
---|---|
Effective start/end date | 5/1/00 → 4/30/06 |
Funding
- National Institute of General Medical Sciences: $235,500.00
- National Institute of General Medical Sciences: $235,500.00
- National Institute of General Medical Sciences: $235,500.00
- National Institute of General Medical Sciences: $46,650.00
- National Institute of General Medical Sciences: $282,150.00
- National Institute of General Medical Sciences: $235,500.00
ASJC
- Genetics
- Molecular Biology
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