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The role of the putative 3′ end processing endonuclease Ysh1p in mRNA and snoRNA synthesis

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posted on 2008-01-01, 00:00 authored by M Garas, Bernhard DichtlBernhard Dichtl, W Keller
Pre-mRNA 3′ end formation is tightly linked to upstream and downstream events of eukaryotic mRNA synthesis. The two-step reaction involves endonucleolytic cleavage of the primary transcript followed by poly(A) addition to the upstream cleavage product. To further characterize the putative 3′ end processing endonuclease Ysh1p/Brr5p, we isolated and analyzed a number of new temperature- and cold-sensitive mutant alleles. We show that Ysh1p plays a crucial role in 3′ end formation and in RNA polymerase II (RNAP II) transcription termination on mRNA genes. In addition, we observed a range of additional functional deficiencies in ysh1 mutant strains, which were partially allele-specific. Interestingly, snoRNA 3′ end formation and RNAP II termination were defective on specific snoRNAs in the cold-sensitive ysh1-12 strain. Moreover, we observed the accumulation of several mRNAs including the NRD1 transcript in this mutant. We provide evidence that NRD1 autoregulation is associated with endonucleolytic cleavage and that this process may involve Ysh1p. In addition, the ysh1-12 strain displayed defects in RNA splicing indicating that a functional link may exist between intron removal and 3′ end formation in yeast. These observations suggest that Ysh1p has multiple roles in RNA synthesis and processing.

History

Journal

RNA

Volume

14

Issue

12

Pagination

2671 - 2684

Publisher

Cold Spring Harbor Laboratory

Location

Woodbury, N.Y.

ISSN

1355-8382

eISSN

1469-9001

Language

eng

Publication classification

C1.1 Refereed article in a scholarly journal

Copyright notice

2008, Cold Spring Harbor Laboratory Press