title: Recognition of distinct RNA motifs by the clustered CCCH zinc fingers of neuronal protein Unkempt.
abstract: Unkempt is an evolutionarily conserved RNA-binding protein that regulates translation of its target genes and is required for the establishment of the early bipolar neuronal morphology. Here we determined the X-ray crystal structure of mouse Unkempt and show that its six CCCH zinc fingers (ZnFs) form two compact clusters, ZnF1-3 and ZnF4-6, that recognize distinct trinucleotide RNA substrates. Both ZnF clusters adopt a similar overall topology and use distinct recognition principles to target specific RNA sequences. Structure-guided point mutations reduce the RNA binding affinity of Unkempt both in vitro and in vivo, ablate Unkempt's translational control and impair the ability of Unkempt to induce a bipolar cellular morphology. Our study unravels a new mode of RNA sequence recognition by clusters of CCCH ZnFs that is critical for post-transcriptional control of neuronal morphology.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.|Division of Newborn Medicine, Boston Children's Hospital, Boston, Massachusetts, USA.
authors.forename: Jernej
authors.initials: J
authors.lastname: Murn
authors.affiliation: Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
authors.forename: Marianna
authors.initials: M
authors.lastname: Teplova
authors.affiliation: Buchmann Institute for Molecular Life Sciences, Goethe University Frankfurt, Frankfurt am Main, Germany.
authors.forename: Kathi
authors.initials: K
authors.lastname: Zarnack
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.|Division of Newborn Medicine, Boston Children's Hospital, Boston, Massachusetts, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
authors.forename: Dinshaw J
authors.initials: DJ
authors.lastname: Patel
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D051379:Mice; D008958:Models, Molecular; D059372:Nucleotide Motifs; D011485:Protein Binding; D011487:Protein Conformation; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D016335:Zinc Fingers
chemical_list: D002352:Carrier Proteins; D016601:RNA-Binding Proteins; C548681:unkempt protein, mouse; D012313:RNA