GENE REGULATION IN RESPONSE
TO ENVIRONMENTAL SIGNALS
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Gisela
Storz, Ph.D., Principal Investigator Orna Carmel-Harel, Ph.D., Postdoctoral Fellow Partha Mukhopadhyay, Ph.D., Postdoctoral Fellow Wayne Outten, Ph.D., Postdoctoral Fellow Xunde Wang, Ph.D., Postdoctoral Fellow Karen Wassarman, Ph.D., Postdoctoral Fellow Matthew Wood, Ph.D., Postdoctoral Fellow Aixia Zhang, Ph.D., Senior Research Assistant David Botstein, Ph.D., Collaborator, Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA Patrick Brown, Ph.D., Collaborator, Biochemistry and Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, CA, USA Susan Gottesman, Ph.D., Collaborator, NCI Robert LaRossa, Ph.D., Collaborator, Biochemical Science and Engineering, E. I. DuPont de Nemours and Company, Wilmington, DE, USA Carsten Rosenow, Ph.D., Collaborator, Affymetrix, Santa Clara, CA, USA Seong-Eon Ryu, Ph.D., Collaborator, Center for Cellular Switch Protein Structure, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon, South Korea Thomas D. Schneider, Ph.D., Collaborator, NCI Ming Zheng, Ph.D., Collaborator, Biochemical Science and Engineering, E. I. DuPont de Nemours and Company, Wilmington, DE, USA |
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The Section on Environmental Gene Regulation studies how Escherichia coli and Saccharomyces cerevisiae cells sense and defend against oxidative stress. Reactive oxygen species can lead to the damage of almost all cell components (DNA, lipid membranes, and proteins) and have been implicated as causative agents in several degenerative diseases. However, most organisms can induce defenses against these oxidants, and it is our goal to understand these adaptive responses. In bacterial cells, a key regulator of the response to hydrogen peroxide is the OxyR transcription factor. OxyR is both the sensor and transducer of the oxidative stress signal; the oxidized but not the reduced form of the purified regulator can activate transcription in vitro. The group previously found that OxyR is activated by the formation of
an intramolecular disulfide bond between C199 and C208 and is deactivated
by enzymatic reduction by glutaredoxin 1 together with glutathione. Recent
structural studies showed that formation of the C199-C208 disulfide bond
leads to a large conformational change. In additional experiments, OxyR
binding sites in the E. coli genome were identified by using a computational
approach, and the transcription profile of the E. coli response to hydrogen
peroxide was determined by microarrays. The chemical basis of OxyR sensitivity
to hydrogen peroxide and the roles of all of OxyR target genes are currently
under investigation. Compared with the bacterial responses to hydrogen
peroxide, little is known about the induction of eukaryotic defenses against
oxidative stress. Thus, the group carried out microarray experiments to
determine genomic expression programs induced by hydrogen peroxide in
wild-type and mutant S. cerevisiae strains. The studies confirmed that
the Yap1 transcription factor is critical for the hydrogen peroxide-dependent
induction of many genes. Genetic screens to isolate mutations in components
of the yeast signal transduction pathways identified thioredoxin reductase
as playing a role in modulating Yap1 activity. Currently, the group is
isolating and characterizing additional mutants and analyzing the purified
Yap1 protein. A second focus of the group is to elucidate the functions of small, untranslated
RNAs. More and more of these RNAs have been shown to play important regulatory
roles. One of the OxyR-induced genes encodes the OxyS RNA, which acts
as a pleiotropic regulator and as an antimutator. OxyS RNA action requires
the Hfq protein, and biochemical experiments have shown that Hfq binds
to the OxyS RNA. This past year, the group found that Hfq is a bacterial
homolog of Sm and Sm-like proteins integral to RNA processing and mRNA
degradation in eukaryotic cells. The nature of the OxyS RNA-Hfq protein
interaction is now being characterized. Programs used to identify protein-encoding
genes generally do not detect small RNA-encoding genes. To try to identify
more of the small RNAs encoded by the E. coli genome, the group used comparative
genomics and microarrays that led to the identification of 17 new small
RNAs, many of which bind to the Hfq protein. Experiments to examine the
global role of Hfq and to elucidate the functions of the newly identified
small RNAs are underway. |
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PUBLICATIONS
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