Processing of a dicistronic small nucleolar RNA precursor by the RNA endonuclease Rnt1.
Small nucleolar RNAs (snoRNAs) are intron encoded or expressed from monocistronic independent transcription units, or, in the case of plants, from polycistronic clusters. We show that the snR190 and U14 snoRNAs from the yeast Saccharomyces cerevisiae are co-transcribed as a dicistronic precursor which is processed by the RNA endonuclease Rnt1, the yeast ortholog of bacterial RNase III. RNT1 disruption results in a dramatic decrease in the levels of mature U14 and snR190 and in accumulation of dicistronic snR190-U14 RNAs. Addition of recombinant Rnt1 to yeast extracts made from RNT1 disruptants induces the chase of dicistronic RNAs into mature snoRNAs, showing that dicistronic RNAs correspond to functional precursors stalled in the processing pathway. Rnt1 cleaves a dicistronic transcript in vitro in the absence of other factors, separating snR190 from U14. Thus, one of the functions of eukaryotic RNase III is, as for the bacterial enzyme, to liberate monocistronic RNAs from polycistronic transcripts.
Chanfreau
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G
Laboratoire du Métabolisme des ARN, URA1300 CNRS, Institut Pasteur, Departement des Biotechnologies, 25 rue du Dr Roux, F-75724 Paris Cedex 15, France. guillom@pasteur.fr
Rotondo
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G
Legrain
P
P
Jacquier
A
A
D001483:Base Sequence; D002874:Chromosome Mapping; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D005796:Genes; D006151:Guanosine; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012315:RNA Caps; D012322:RNA Precursors / Q000737:chemistry; D012323:RNA Processing, Post-Transcriptional; D012331:RNA, Fungal / Q000737:chemistry; D012342:RNA, Small Nuclear / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000235:genetics; D043244:Ribonuclease III; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins
D012315:RNA Caps; D012322:RNA Precursors; D012331:RNA, Fungal; D012342:RNA, Small Nuclear; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D006151:Guanosine; D004722:Endoribonucleases; C468367:RNT1 protein, S cerevisiae; D043244:Ribonuclease III