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Molecular and Cellular Biology, February 2004, p. 945-953, Vol. 24, No. 3
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.3.945-953.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

RNase MRP Cleaves the CLB2 mRNA To Promote Cell Cycle Progression: Novel Method of mRNA Degradation

Tina Gill, Ti Cai, Jason Aulds, Sara Wierzbicki, and Mark E. Schmitt*

Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, Syracuse, New York 13210

Received 24 October 2003/ Accepted 3 November 2003

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.


* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, 750 East Adams St., Syracuse, NY 13210. Phone: (315) 464-8713. Fax: (315) 464-8750. E-mail: schmittm{at}upstate.edu.


Molecular and Cellular Biology, February 2004, p. 945-953, Vol. 24, No. 3
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.3.945-953.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.




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