title: RNA Editing TUTase 1: structural foundation of substrate recognition, complex interactions and drug targeting.
abstract: Terminal uridyltransferases (TUTases) execute 3' RNA uridylation across protists, fungi, metazoan and plant species. Uridylation plays a particularly prominent role in RNA processing pathways of kinetoplastid protists typified by the causative agent of African sleeping sickness, Trypanosoma brucei In mitochondria of this pathogen, most mRNAs are internally modified by U-insertion/deletion editing while guide RNAs and rRNAs are U-tailed. The founding member of TUTase family, RNA editing TUTase 1 (RET1), functions as a subunit of the 3' processome in uridylation of gRNA precursors and mature guide RNAs. Along with KPAP1 poly(A) polymerase, RET1 also participates in mRNA translational activation. RET1 is divergent from human TUTases and is essential for parasite viability in the mammalian host and the insect vector. Given its robust in vitro activity, RET1 represents an attractive target for trypanocide development. Here, we report high-resolution crystal structures of the RET1 catalytic core alone and in complex with UTP analogs. These structures reveal a tight docking of the conserved nucleotidyl transferase bi-domain module with a RET1-specific C2H2 zinc finger and RNA recognition (RRM) domains. Furthermore, we define RET1 region required for incorporation into the 3' processome, determinants for RNA binding, subunit oligomerization and processive UTP incorporation, and predict druggable pockets.
authors.affiliation: Department of Molecular Biology, University of Geneva, 1211 Geneva, Switzerland.
authors.forename: Lional
authors.initials: L
authors.lastname: Rajappa-Titu
authors.affiliation: Department of Molecular and Cell Biology, Boston University School of Dental Medicine, Boston, MA 02118, USA.
authors.forename: Takuma
authors.initials: T
authors.lastname: Suematsu
authors.affiliation: Department of Molecular Biology, University of Geneva, 1211 Geneva, Switzerland.
authors.forename: Paola
authors.initials: P
authors.lastname: Munoz-Tello
authors.affiliation: Department of Molecular Biology, University of Geneva, 1211 Geneva, Switzerland.
authors.forename: Marius
authors.initials: M
authors.lastname: Long
authors.affiliation: Department of Chemistry & Biochemistry and the National Biomedical Computation Resource, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Özlem
authors.initials: Ö
authors.lastname: Demir
authors.affiliation: Department of Chemistry & Biochemistry and the National Biomedical Computation Resource, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Cheng
authors.affiliation: Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697, USA.
authors.forename: Jason R
authors.initials: JR
authors.lastname: Stagno
authors.affiliation: Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697, USA.
authors.forename: Hartmut
authors.initials: H
authors.lastname: Luecke
authors.affiliation: Department of Chemistry & Biochemistry and the National Biomedical Computation Resource, University of California, San Diego, La Jolla, CA 92093, USA.
authors.forename: Rommie E
authors.initials: RE
authors.lastname: Amaro
authors.affiliation: Department of Molecular and Cell Biology, Boston University School of Dental Medicine, Boston, MA 02118, USA.
authors.forename: Inna
authors.initials: I
authors.lastname: Aphasizheva
authors.affiliation: Department of Molecular and Cell Biology, Boston University School of Dental Medicine, Boston, MA 02118, USA ruslana@bu.edu.|Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Ruslan
authors.initials: R
authors.lastname: Aphasizhev
authors.affiliation: Department of Molecular Biology, University of Geneva, 1211 Geneva, Switzerland stephane.thore@inserm.fr.|INSERM, U1212, ARNA Laboratory, Bordeaux 33000, France.|CNRS UMR5320, ARNA Laboratory, Bordeaux 33000, France.|University of Bordeaux, ARNA Laboratory, Bordeaux 33000, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Thore
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D020756:Coatomer Protein / Q000737:chemistry*; D018360:Crystallography, X-Ray; D015195:Drug Design; D006860:Hydrogen Bonding; D007700:Kinetics; D007891:Leishmania / Q000201:enzymology; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D015800:Protozoan Proteins / Q000737:chemistry*; D017393:RNA Editing; D013379:Substrate Specificity; D014344:Trypanocidal Agents / Q000737:chemistry
chemical_list: D020756:Coatomer Protein; D015800:Protozoan Proteins; D014344:Trypanocidal Agents