Seven novel methylation guide small nucleolar RNAs are processed from a common polycistronic transcript by Rat1p and RNase III in yeast.
Through a computer search of the genome of the yeast Saccharomyces cerevisiae, the coding sequences of seven different box C/D antisense small nucleolar RNAs (snoRNAs) with the structural hallmarks of guides for rRNA ribose methylation have been detected clustered over a 1.4-kb tract in an inter-open reading frame region of chromosome XIII. The corresponding snoRNAs have been positively identified in yeast cells. Disruption of the nonessential snoRNA gene cluster specifically suppressed the seven cognate rRNA ribose methylations but did not result in any growth delay under the conditions of yeast culture tested. The seven snoRNAs are processed from a common polycistronic transcript synthesized from an independent promoter, similar to some plant snoRNAs but in marked contrast with their vertebrate functional homologues processed from pre-mRNA introns containing a single snoRNA. Processing of the polycistronic precursor requires nucleases also involved in rRNA processing, i.e., Rnt1p and Rat1p. After disruption of the RNT1 gene, the yeast ortholog of bacterial RNase III, production of the seven mature snoRNAs was abolished, while the polycistronic snoRNA precursor accumulated. In cells lacking functional Rat1p, an exonuclease involved in the processing of both pre-rRNA and intron-encoded snoRNAs, several processing intermediates of the polycistronic precursor accumulated. This allowed for the mapping in the precursor of the presumptive Rnt1p endonucleolytic cuts which provide entry sites for subsequent exonucleolytic trimming of the pre-snoRNAs. In line with known properties of double-stranded RNA-specific RNase III, pairs of Rnt1p cuts map next to each other on opposite strands of long double-helical stems in the secondary structure predicted for the polycistronic snoRNA precursor.
Qu
L H
LH
Biotechnology Research Center, Zhongshan University, Guangzhou 510 275, People's Republic of China.
Henras
A
A
Lu
Y J
YJ
Zhou
H
H
Zhou
W X
WX
Zhu
Y Q
YQ
Zhao
J
J
Henry
Y
Y
Caizergues-Ferrer
M
M
Bachellerie
J P
JP
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004722:Endoribonucleases / Q000378:metabolism*; D005095:Exoribonucleases / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D015870:Gene Expression; D005800:Genes, Fungal; D008745:Methylation; D008969:Molecular Sequence Data; D005810:Multigene Family; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D012323:RNA Processing, Post-Transcriptional; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000096:biosynthesis; D012342:RNA, Small Nuclear / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D043244:Ribonuclease III; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D058727:RNA, Small Untranslated
D017931:DNA Primers; D004271:DNA, Fungal; D005656:Fungal Proteins; D016372:RNA, Antisense; D012331:RNA, Fungal; D012335:RNA, Ribosomal; D012342:RNA, Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; C075577:RAT1 protein, S cerevisiae; D004722:Endoribonucleases; D005095:Exoribonucleases; C468367:RNT1 protein, S cerevisiae; D043244:Ribonuclease III; D058727:RNA, Small Untranslated