Analysis of CUGBP1 targets identifies GU-repeat sequences that mediate rapid mRNA decay.
CUG-repeat binding protein 1 (CUGBP1) mediates selective mRNA decay by binding to GU-rich elements (GREs) containing the sequence UGUUUGUUUGU found in the 3' untranslated region (UTR) of short-lived transcripts. We used an anti-CUGBP1 antibody to immunoprecipitate CUGBP1 from HeLa cytoplasmic extracts and analyzed the associated transcripts using oligonucleotide microarrays. We identified 613 putative mRNA targets of CUGBP1 and found that the UGUUUGUUUGU GRE sequence and a GU-repeat sequence were both highly enriched in the 3' UTRs of these targets. We showed that CUGBP1 bound specifically to the GU-repeat sequence and that insertion of this sequence into the 3' UTR of a beta-globin reporter transcript conferred instability to the transcript. Based on these results, we redefined the GRE to include this GU-repeat sequence. Our results suggest that CUGBP1 coordinately regulates the mRNA decay of a network of transcripts involved in cell growth, cell motility, and apoptosis.
Rattenbacher
Bernd
B
Center for Infectious Diseases and Microbiology Translational Research, Department of Microbiology, University of Minnesota, Minneapolis, MN 55455, USA.
Beisang
Daniel
D
Wiesner
Darin L
DL
Jeschke
Jonathan C
JC
von Hohenberg
Maximilian
M
St Louis-Vlasova
Irina A
IA
Bohjanen
Paul R
PR
D020413:3' Untranslated Regions; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D000067879:CELF1 Protein; D016384:Consensus Sequence; D015226:Dinucleoside Phosphates / Q000378:metabolism; D018900:Dinucleotide Repeats; D055785:Gene Knockdown Techniques; D053263:Gene Regulatory Networks; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D011485:Protein Binding; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
D020413:3' Untranslated Regions; D000067879:CELF1 Protein; C102843:CELF1 protein, human; D015226:Dinucleoside Phosphates; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; C018310:guanylyl-(3'-5')-uridine