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Steven J. Brill
Professor
Rutgers University
Dept. of Molecular Biology and Biochemistry
CABM - 679 Hoes Lane
Piscataway. NJ 08854
(732) 235-4197
FAX - 4880
brill@cabm.rutgers.edu |
DNA replication. DNA repair. DNA
helicase. genetic analysis. genome
stability. protein purification. yeast
Our lab uses the
budding yeast S. cerevisiae as a model system to study DNA replication
and genome stability in eukaryotic cells. Our approach is to apply genetics. biochemistry and molecular biology to analyze highly-conserved genes involved
in this process. The main project focuses on a DNA helicase-topoisomerase
complex that is known to control genome stability in humans. The yeast gene SGS1 is the homolog of the genes responsible for both Bloom's Syndrome
and Werner's Syndrome in humans. We have shown that Sgs1 is a 3' - 5' DNA
helicase that interacts with Top3 and is composed of two functional domains. In the absence of either domain the yeast genome undergoes hyper-recombination
and rearrangement. A second project involves the use of genetic screens to
identify and characterize new genes required for genome stability. Here we
are focused on determining the function of six conserved but poorly
characterized "SLX" genes that were identified based on their genetic
interaction with SGS1. Enzymatically. we have shown that four of these
these genes encode two heterodimeric structure-specific endonucleases while a
third pair appears to encode a novel protein-modifying activity. The
long-term goal of this project is to determine the mechanism by which these
enzymes maintain genome stability. A third project involves Replication
Protein A (RPA) which is a ssDNA binding protein required for DNA replication. repair. and recombination. RPA is a three-subunit ssDNA binding protein that
is found in all eukaryotes from yeast to humans. Using a combination of
biochemistry and molecular biology we have shown that RPA contains at least
four non-identical ssDNA binding domains. Genetically. we have determined
which of these domains are required for cell viability. and have identified
other essential regions of the protein that interact directly with the DNA
replication machinery. Our current focus is to determine the role of RPA in
the DNA damage response.
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