RNase MRP cleaves the CLB2 mRNA to promote cell cycle progression: novel method of mRNA degradation.
RNase mitochondrial RNA processing (RNase MRP) mutants have been shown to have an exit-from-mitosis defect that is caused by an increase in CLB2 mRNA levels, leading to increased Clb2p (B-cyclin) levels and a resulting late anaphase delay. Here we describe the molecular defect behind this delay. CLB2 mRNA normally disappears rapidly as cells complete mitosis, but the level remains high in RNase MRP mutants. This is in direct contrast to other exit-from-mitosis mutants and is the result of an increase in CLB2 mRNA stability. We found that highly purified RNase MRP cleaved the 5' untranslated region (UTR) of the CLB2 mRNA in several places in an in vitro assay. In vivo, we identified RNase MRP-dependent cleavage products on the CLB2 mRNA that closely matched in vitro products. Disposal of these products was dependent on the 5'-->3' exoribonuclease Xrn1 and not the exosome. Our results demonstrate that the endoribonuclease RNase MRP specifically cleaves the CLB2 mRNA in its 5'-UTR to allow rapid 5' to 3' degradation by the Xrn1 nuclease. Degradation of the CLB2 mRNA by the RNase MRP endonuclease provides a novel way to regulate the cell cycle that complements the protein degradation machinery. In addition, these results denote a new mechanism of mRNA degradation not seen before in the yeast Saccharomyces cerevisiae.
Gill
Tina
T
Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, Syracuse, New York 13210, USA.
Cai
Ti
T
Aulds
Jason
J
Wierzbicki
Sara
S
Schmitt
Mark E
ME
D020121:5' Untranslated Regions / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D002453:Cell Cycle / Q000502:physiology*; D019926:Cyclin B / Q000096:biosynthesis / Q000235:genetics*; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D005095:Exoribonucleases / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
D020121:5' Untranslated Regions; C093749:CLB2 protein, S cerevisiae; D019926:Cyclin B; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D004722:Endoribonucleases; D005095:Exoribonucleases; C058330:mitochondrial RNA-processing endoribonuclease; C068398:XRN1 protein, S cerevisiae