Rainer B. Lanz, Bahram Razani, Aaron D. Goldberg, and Bert W. O'Malley*
Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030
* To whom correspondence should be addressed. E-mail: berto@bcm.tmc.edu
Steroid receptor RNA activator (SRA) is an RNA transcript that functions
as a eukaryotic transcriptional coactivator for steroid hormone receptors.
We report here the isolation and functional characterization of distinct
RNA substructures within the SRA molecule that constitute its coactivation
function. We used comparative sequence analysis and free energy calculations
to systematically study SRA RNA subdomains for identification of structured
regions and base pairings, and we used site-directed mutagenesis to assess
their functional consequences. Together with genetic deletion analysis,
this approach identified six RNA motifs in SRA important for coactivation.
Because all nucleotide changes in the mutants that disrupted SRA function
were silent mutations presumed not to alter deduced encoded amino acid
sequence, our analysis provides strong evidence that SRA-mediated coactivation
is executed by distinct RNA motifs and not by an encoded
protein.
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