Systematic genomic screening and analysis of mRNA in untranslated regions and mRNA precursors: combining experimental and computational approaches.
BACKGROUND
The untranslated regions (UTRs) of mRNA upstream (5'UTR) and downstream (3'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'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'UTR of the estrogen receptor mRNA in this way.


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


BACKGROUND
dandekar@embl-heidelberg.de
Dandekar
T
T
European Molecular Biology Laboratory, Postfach 102209, D-69012 Heidelberg, Germany. dandekar@embl-heidelberg.de
Beyer
K
K
Bork
P
P
Kenealy
M R
MR
Pantopoulos
K
K
Hentze
M
M
Sonntag-Buck
V
V
Flouriot
G
G
Gannon
F
F
Schreiber
S
S
D020413:3' Untranslated Regions / Q000235:genetics; D002874:Chromosome Mapping / Q000379:methods; D019295:Computational Biology / Q000379:methods*; D004926:Escherichia coli; D006801:Humans; D007501:Iron / Q000378:metabolism; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods*; D012986:Software Validation
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D007501:Iron