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Biochim Biophys Acta. 2009 Jun-Aug;1789(6-8):469-76. doi: 10.1016/j.bbagrm.2009.05.003. Epub 2009 May 19.

Compensatory mutations in the L30e kink-turn RNA-protein complex.

Author information

  • 1Department of Chemistry, Bryn Mawr College, Bryn Mawr, PA 19010, USA.

Abstract

The S. cerevisiae ribosomal protein L30e is an autoregulatory protein that binds to its own pre-mRNA and mature mRNA to inhibit splicing and translation, respectively. The L30e RNA-binding element is a stem-asymmetric loop-stem that forms a kink-turn. A bacterial genetic system was designed to test the ability of protein variants to repress the expression of reporter mRNAs containing the L30e RNA-binding element. Initial screens revealed that changes in several RNA nucleotides had a measurable effect on repression of the reporter by the wild type protein. RNA mutants that reduce repression were screened against libraries of randomly mutagenized L30e proteins. These screens identified a glycine to serine mutation of L30e, which specifically restores activity to an RNA variant containing a U that replaces a helix-capping G. Similarly, an asparagine to alanine mutation was found to suppress a substitution at a position where the L30e RNA nucleotide extends out into the protein pocket. In addition, a compensatory RNA mutation within a defective RNA variant was found. The identification of these suppressors provides new insights into the architecture of a functional binding element and its recognition by an important RNA-binding protein.

PMID:
19460470
[PubMed - indexed for MEDLINE]
PMCID:
PMC2743985
Free PMC Article

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