title: A novel cis-acting element from the 3'UTR of DNA damage-binding protein 2 mRNA links transcriptional and post-transcriptional regulation of gene expression.
abstract: The DNA damage-binding protein 2 (DDB2) is an adapter protein that can direct a modular Cul4-DDB1-RING E3 Ligase complex to sites of ultraviolet light-induced DNA damage to ubiquitinate substrates during nucleotide excision repair. The DDB2 transcript is ultraviolet-inducible; therefore, its regulation is likely important for its function. Curiously, the DDB2 mRNA is reportedly short-lived, but the transcript does not contain any previously characterized cis-acting determinants of mRNA stability in its 3' untranslated region (3'UTR). Here, we used a tetracycline regulated d2EGFP reporter construct containing specific 3'UTR sequences from DDB2 to identify novel cis-acting elements that regulate mRNA stability. Synthetic 3'UTRs corresponding to sequences as short as 25 nucleotides from the central region of the 3'UTR of DDB2 were sufficient to accelerate decay of the heterologous reporter mRNA. Conversely, these same 3'UTRs led to more rapid induction of the reporter mRNA, export of the message to the cytoplasm and the subsequent accumulation of the encoded reporter protein, indicating that this newly identified cis-acting element affects transcriptional and post-transciptional processes. These results provide clear evidence that nuclear and cytoplasmic processing of the DDB2 mRNA is inextricably linked.
authors.affiliation: Cancer Therapeutics Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada, K1H 8L6.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Melanson
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Cabrita
authors.forename: Reetesh
authors.initials: R
authors.lastname: Bose
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Hamill
authors.forename: Elysia
authors.initials: E
authors.lastname: Pan
authors.forename: Christian
authors.initials: C
authors.lastname: Brochu
authors.forename: Kristen A
authors.initials: KA
authors.lastname: Marcellus
authors.forename: Tong T
authors.initials: TT
authors.lastname: Zhao
authors.forename: Martin
authors.initials: M
authors.lastname: Holcik
authors.forename: Bruce C
authors.initials: BC
authors.lastname: McKay
mesh_terms: D020413:3' Untranslated Regions; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D055029:Inverted Repeat Sequences; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D038621:Regulatory Sequences, Ribonucleic Acid; D014158:Transcription, Genetic
chemical_list: D020413:3' Untranslated Regions; C117669:DDB2 protein, human; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D038621:Regulatory Sequences, Ribonucleic Acid; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins