Systematic Genomic Screening and Analysis of mRNA in Untranslated Regions and mRNA Precursors: Combining Experimental and Computational Approaches

Thomas Dandekar1,2, Katrin Beyer3, Peer Bork1,2, Mary-Rose Kenealy1, Kostas Pantopoulos1, Mathias Hentze1, Vera Sonntag-Buck1, Gilles Flouriot1, Frank Gannon 1 and Sonja Schreiber1

1 European Molecular Biology Laboratory, Postfach 102209, D-69012 Heidelberg,
2 Max-Delbrück-Centrum für Molekulare Medizin, Robert-Rösslestr. 10, 13122 Berlin, Germany and
3 Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland

Bioinformatics, 14(3), 271-278 (April 1998)

Abstract

Motivation: The untranslated regions (UTRs) of mRNA upstream (5[prime]UTR) and downstream (3[prime]UTR) of the open reading frame, as well as the mRNA precursor, carry important regulatory sequences. To reveal unidentified regulatory signals, we combine information from experiments with computational approaches. Depending on available knowledge, three different strategies are employed.

Results: Searching with a consensus template, new RNAs with regulatory RNA elements can be identified in genomic screens. By this approach, we identify new candidate regulatory motifs resembling iron-responsive elements in the 5[prime]UTRs of HemA, FepB and FrdB mRNA from Escherichia coli. If an RNA element is not yet defined, it may be analyzed by combining results from SELEX (selective enrichment of ligands by exponential amplification) and a search of databases from RNA or genomic sequences. A cleavage stimulating factor (CstF) binding element 3 of the polyadenylation site in the mRNA precursor serves as a test example. Alternatively, the regulatory RNA element may be found by studying different RNA foldings and their correlation with simple experimental tests. We delineate a novel instability element in the 3[prime]UTR of the estrogen receptor mRNA in this way.

Availability: Strategy, methods and programs are available on request from T.Dandekar.

Contact: dandekar@embl-heidelberg.de