Regulation of the Ysh1 endonuclease of the mRNA cleavage/polyadenylation complex by ubiquitin-mediated degradation

Mutation of the essential yeast protein Ipa1 has previously been demonstrated to cause defects in pre-mRNA 3ʹ end processing and growth, but the mechanism underlying these defects was not clear. In this study, we show that the <i>ipa1-1</i> mutation causes a striking depletion of Ysh1, the evolutionarily conserved endonuclease subunit of the 19-subunit mRNA Cleavage/Polyadenylation (C/P) complex, but does not decrease other C/P subunits. <i>YSH1</i> overexpression rescues both the growth and 3ʹ end processing defects of the <i>ipa1-1</i> mutant. <i>YSH1</i> mRNA level is unchanged in <i>ipa1-1</i> cells, and proteasome inactivation prevents Ysh1 loss and causes accumulation of ubiquitinated Ysh1. Ysh1 ubiquitination is mediated by the Ubc4 ubiquitin-conjugating enzyme and Mpe1, which in addition to its function in C/P, is also a RING ubiquitin ligase. In summary, Ipa1 affects mRNA processing by controlling the availability of the C/P endonuclease and may represent a regulatory mechanism that could be rapidly deployed to facilitate reprogramming of cellular responses.