Abstract
Exoribonucleases function in the processing and degradation of a variety of RNAs in all organisms. These enzymes play a particularly important role in the maturation of rRNAs and in a quality-control pathway that degrades rRNA precursors upon inhibition of ribosome biogenesis. Strains with defects in 3'-5' exoribonucleolytic components of the RNA processing exosome accumulate polyadenylated precursor rRNAs that also arise in strains with ribosome biogenesis defects. These findings suggested that polyadenylation might target pre-rRNAs for degradation by the exosome. Here we report experiments that indicate a role for the 5'-3' exoribonuclease Rat1p and its associated protein Rai1p in the degradation of poly(A)(+) pre-rRNAs. Depletion of Rat1p enhances the amount of poly(A)(+) pre-rRNA that accumulates in strains deleted for the exosome subunit Rrp6p and decreases their 5' heterogeneity. Deletion of RAI1 results in the accumulation of poly(A)(+) pre-rRNAs, and inhibits Rat1p-dependent 5'-end processing and Rrp6p-dependent 3'-end processing of 5.8S rRNA. RAT1 and RAI1 mutations cause synergistic growth defects in the presence of rrp6-Delta, consistent with the interdependence of 5'-end and 3'-end processing pathways. These findings suggest that Rai1p may coordinate the 5'-end and 3'-end processing and degradation activities of Rat1p and the nuclear exosome.
MeSH Terms
Antimetabolites/pharmacology
Exoribonucleases/genetics,metabolism
Fluorouracil/pharmacology
Fungal Proteins/metabolism
Models, Biological
Nuclear Proteins/genetics,metabolism
RNA Precursors/metabolism
RNA Processing, Post-Transcriptional
RNA, Fungal/genetics,metabolism
RNA, Ribosomal/metabolism
RNA-Binding Proteins
Reverse Transcriptase Polymerase Chain Reaction
Saccharomyces cerevisiae/enzymology,genetics,growth & development,metabolism
Saccharomyces cerevisiae Proteins/genetics,metabolism
Chemicals
Antimetabolites
Fungal Proteins
Nuclear Proteins
RNA Precursors
RNA, Fungal
RNA, Ribosomal
RNA-Binding Proteins
Rai1 protein, S cerevisiae
Saccharomyces cerevisiae Proteins
RAT1 protein, S cerevisiae
Exoribonucleases
Fluorouracil
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fang Feng
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA.
Phillips Seasson
Butler J Scott
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