title: Conserved binding of GCAC motifs by MEC-8, couch potato, and the RBPMS protein family.
abstract: Precise regulation of mRNA processing, translation, localization, and stability relies on specific interactions with RNA-binding proteins whose biological function and target preference are dictated by their preferred RNA motifs. The RBPMS family of RNA-binding proteins is defined by a conserved RNA recognition motif (RRM) domain found in metazoan RBPMS/Hermes and RBPMS2,Drosophilacouch potato, and MEC-8 fromCaenorhabditis elegansIn order to determine the parameters of RNA sequence recognition by the RBPMS family, we have first used the N-terminal domain from MEC-8 in binding assays and have demonstrated a preference for two GCAC motifs optimally separated by >6 nucleotides (nt). We have also determined the crystal structure of the dimeric N-terminal RRM domain from MEC-8 in the unbound form, and in complex with an oligonucleotide harboring two copies of the optimal GCAC motif. The atomic details reveal the molecular network that provides specificity to all four bases in the motif, including multiple hydrogen bonds to the initial guanine. Further studies with human RBPMS, as well asDrosophilacouch potato, confirm a general preference for this double GCAC motif by other members of the protein family and the presence of this motif in known targets.
authors.affiliation: University of Bordeaux, Institut Européen de Chimie et Biologie, F-33607 Pessac, France.|Inserm U1212, CNRS UMR 5320, ARNA Laboratory, F-33076 Bordeaux, France.
authors.forename: Heddy
authors.initials: H
authors.lastname: Soufari
authors.affiliation: University of Bordeaux, Institut Européen de Chimie et Biologie, F-33607 Pessac, France.|Inserm U1212, CNRS UMR 5320, ARNA Laboratory, F-33076 Bordeaux, France.
authors.forename: Cameron D
authors.identifier: 0000-0002-0776-7947
authors.initials: CD
authors.lastname: Mackereth
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D006860:Hydrogen Bonding; D008958:Models, Molecular; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D059372:Nucleotide Motifs; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D029742:Caenorhabditis elegans Proteins; D029721:Drosophila Proteins; D009687:Nuclear Proteins; C101827:RBPMS protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C078019:cpo protein, Drosophila; C099356:mec-8 protein, C elegans
keywords: ITC; NMR; X-ray crystallography; protein–RNA; structure