Abstract
Publisher Summary There are potentially important differences between the activity and intracellular locations of XRNs of yeast and multicellular eukaryotes. This chapter describes experiments that make use of complementation of yeast xrn1 and xrn2/rat1 mutants in combination with poly(G) tract studies, which allow for these differences to be addressed. These enzymes are similar in sequence and enzymatic activity but differ in their intracellular locations and cellular functions. Xrn1p is an abundant cytoplasmic enzyme and plays a major role in the degradation of mRNAs in the cytoplasm, as well as trimming the 5′ ends of rRNAs and degrading rRNA processing intermediates. In contrast to Xrn1p, Xrn2p/Rat1p is targeted to the yeast nucleus, exists in a complex with Rai1p, and functions in the processing of rRNA and small nucleolar RNAs (snoRNAs). The results of these studies can yield important insight into the possible function of XRN enzymes and can be used in conjunction with more detailed analysis of the XRN enzymes in their native contexts.
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