An AU-rich element in the 3' untranslated region of the C/EBP delta mRNA is important for protein binding during G0 growth arrest.
Posttranscriptional regulation at the level of mRNA stability is becoming increasingly recognized as an important mechanism to control the levels of mRNAs that encode key cell fate determining proteins. Previous work from our laboratory demonstrated that C/EBPdelta is a highly unstable mRNA in G(0) growth arrested mammary epithelial cells. In this report we investigated trans-acting factor binding to the C/EBPdelta 3'-UTR and identified a cis-acting element important for this interaction. RNA electromobility shift assays (REMSAs) demonstrate that the C/EBPdelta mRNA 3'-UTR binds trans-acting factor(s) present in G(0) growth arrested mammary epithelial cell lysates. This binding was not detected in the presence of lysates from growing cells. UV-binding analysis detected a RNA/protein complex of approximately 35kDa following incubation of the full-length C/EBPdelta 3'UTR with lysates from G(0) growth arrested mammary epithelial cells. Competition assays indicate that a specific AU-rich region (U1) is necessary for trans-acting factor binding to the C/EBPdelta 3'-UTR. These studies have identified an AU-rich element located within the C/EBPdelta 3'-UTR that interacts with a putative G(0) growth arrest-specific trans-acting factor(s), which may regulate C/EBPdelta mRNA decay.
Dearth
Lawrence R
LR
Molecular, Cellular and Developmental Biology Graduate Program, The Ohio State University, Columbus, OH 43210-1093, USA.
DeWille
James
J
D020413:3' Untranslated Regions / Q000737:chemistry* / Q000378:metabolism*; D000241:Adenosine / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D051743:CCAAT-Enhancer-Binding Protein-delta; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics*; D002455:Cell Division; D002460:Cell Line; D024202:Electrophoretic Mobility Shift Assay; D051379:Mice; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D020871:RNA Stability; D016601:RNA-Binding Proteins / Q000378:metabolism*; D038621:Regulatory Sequences, Ribonucleic Acid; D016192:Resting Phase, Cell Cycle; D014157:Transcription Factors; D014529:Uridine / Q000032:analysis
D020413:3' Untranslated Regions; D022762:CCAAT-Enhancer-Binding Proteins; C497253:Cebpd protein, mouse; D016601:RNA-Binding Proteins; D038621:Regulatory Sequences, Ribonucleic Acid; D014157:Transcription Factors; D051743:CCAAT-Enhancer-Binding Protein-delta; D000241:Adenosine; D014529:Uridine