title: Antisense Transcripts Delimit Exonucleolytic Activity of the Mitochondrial 3' Processome to Generate Guide RNAs.
abstract: Small, noncoding RNA biogenesis typically involves cleavage of structured precursor by RNase III-like endonucleases. However, guide RNAs (gRNAs) that direct U-insertion/deletion mRNA editing in mitochondria of trypanosomes maintain 5' triphosphate characteristic of the transcription initiation and possess a U-tail indicative of 3' processing and uridylation. Here, we identified a protein complex composed of RET1 TUTase, DSS1 3'-5' exonuclease, and three additional subunits. This complex, termed mitochondrial 3' processome (MPsome), is responsible for primary uridylation of ∼800 nt gRNA precursors, their processive degradation to a mature size of 40-60 nt, and secondary U-tail addition. Both strands of the gRNA gene are transcribed into sense and antisense precursors of similar lengths. Head-to-head hybridization of these transcripts blocks symmetrical 3'-5' degradation at a fixed distance from the double-stranded region. Together, our findings suggest a model in which gRNA is derived from the 5' extremity of a primary molecule by uridylation-induced, antisense transcription-controlled 3'-5' exonucleolytic degradation.
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: Section of Computational Biomedicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Liye
authors.initials: L
authors.lastname: Zhang
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: Section of Computational Biomedicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Stefano
authors.initials: S
authors.lastname: Monti
authors.affiliation: Department of Physiology & Biophysics, School of Medicine, University of California, Irvine, Irvine, CA 92697, USA.
authors.forename: Lan
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Costello
authors.affiliation: Department of Molecular and Cell Biology, Boston University School of Dental Medicine, Boston, MA 02118, USA; Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA. Electronic address: ruslana@bu.edu.
authors.forename: Ruslan
authors.initials: R
authors.lastname: Aphasizhev
mesh_terms: D005095:Exoribonucleases / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D008928:Mitochondria / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis* / Q000235:genetics; D017393:RNA Editing; D012316:RNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism*; D017394:RNA, Guide, Kinetoplastida / Q000096:biosynthesis* / Q000235:genetics; D000077278:RNA, Mitochondrial; D016053:RNA, Protozoan / Q000096:biosynthesis* / Q000235:genetics; D013997:Time Factors; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000378:metabolism*; D014500:Uracil Nucleotides / Q000378:metabolism
chemical_list: D015800:Protozoan Proteins; D016372:RNA, Antisense; D000077278:RNA, Mitochondrial; D016053:RNA, Protozoan; D014500:Uracil Nucleotides; D012313:RNA; D012316:RNA Nucleotidyltransferases; D005095:Exoribonucleases; D017394:RNA, Guide, Kinetoplastida
keywords: RNA decay; RNA editing; TUTase; exonuclease; guide RNA; mitochondria; trypanosoma
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