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Brenda Bass

Professor of Biochemistry

B.A. Colorado College

Ph.D. University of Colorado, Boulder

Research

References

 

Brenda Bass's Lab Page

Research

Research in my laboratory is focused on double-stranded RNA (dsRNA)--its biologic functions and the proteins that bind it to mediate these functions. Our studies are divided between two dsRNA-mediated pathways: RNA editing by adenosine deaminases that act on RNA (ADARs), and RNA interference (RNAi). For both pathways we perform in vitro studies to answer mechanistic questions, and in vivo studies in C. elegans to understand biologic function. dsRNA binding proteins (dsRBPs) bind tightly to dsRNA of any sequence, and a dsRNA substrate for one dsRBP is also a substrate for others. This suggests that different dsRNA-mediated pathways intersect and affect each other, and thus, we also study how RNA editing affects RNAi.

ADARs deaminate adenosines in double-stranded regions of cellular and viral RNAs to create the nucleoside inosine. Inosine is read as guanosine by the translational machinery, and one function of ADARs is to deaminate adenosines within codons, so that multiple protein isoforms can be synthesized from a single, encoded mRNA. In this way ADARs create protein isoforms of hepatitis delta antigen, and many proteins involved in neurotransmission, such as serotonin receptors and glutamate receptors. Consistent with the observation that ADARs are highly expressed in the nervous system, mutant animals lacking ADARs have behavioral defects. For example, our characterizations of C. elegans strains lacking ADARs show that these animals have aberrant chemotaxis and thermotaxis behaviors.

Several years ago my laboratory developed a method to identify inosine-containing RNA, and applied the method to mRNA isolated from C. elegans and human brain. We found inosines in remarkably stable structures that sometimes contain hundreds of nearly contiguous base-pairs. Surprisingly, the edited structures were in non-coding regions of mRNAs, such as 5' and 3' untranslated regions (UTRs), and introns. Our data suggest non-coding sequences are the primary targets of ADARs and that ADARs have functions in addition to altering codon meaning. Researchers in my lab are attempting to understand the function of these long double-stranded structures and the inosines within them.

In collaboration with Chris Hill's lab (University of Utah), we solved the crystal structure of the catalytic domain of human ADAR2. This study shows that ADAR catalysis involves a catalytic zinc coordinated to two cysteines and a histidine, as predicted from sequence similarities to the cytidine deaminase family. Current biochemical studies are focused on testing hypotheses about mechanism, substrate recognition, and substrate specificity that are suggested by the structure.

In addition to our studies on how RNA editing affects RNAi, we are also interested in the RNAi pathway in its own right, in particular, as it occurs in C. elegans. Biochemical studies are focused on the dsRBPs required for RNAi in C. elegans, and we analyze their RNA binding properties and their catalytic activities in vitro. In regard to biologic function we want to understand the natural roles of dsRNA-mediated gene silencing. To this end we have used microarray analyses to identify genes that are misregulated in C. elegans strains containing mutations in the Dicer gene, as well as those with mutations in other genes required for RNAi.

References