Poly(A) tail-dependent exonuclease AtRrp41p from Arabidopsis thaliana rescues 5.8 S rRNA processing and mRNA decay defects of the yeast ski6 mutant and is found in an exosome-sized complex in plant and yeast cells.
Eukaryotic 3'-->5' exonucleolytic activities are essential for a wide variety of reactions of RNA maturation and metabolism, including processing of rRNA, small nuclear RNA, and small nucleolar RNA, and mRNA decay. Two related but distinct forms of a complex containing 10 3'-->5' exonucleases, the exosome, are found in yeast nucleus and cytoplasm, respectively, and related complexes exist in human cells. Here we report on the characterization of the AtRrp41p, an Arabidopsis thaliana homolog of the Saccharomyces cerevisiae exosome subunit Rrp41p (Ski6p). Purified recombinant AtRrp41p displays a processive phosphorolytic exonuclease activity and requires a single-stranded poly(A) tail on a substrate RNA as a "loading pad." The expression of the Arabidopsis RRP41 cDNA in yeast rescues the 5.8 S rRNA processing and 3'-->5' mRNA degradation defects of the yeast ski6-100 mutant. However, neither of these defects can explain the conditional lethal phenotype of the ski6-100 strain. Importantly, AtRrp41p shares additional function(s) with the yeast Rrp41p which are essential for cell viability because it also rescues the rrp41 (ski6) null mutant. AtRrp41p is found predominantly in a high molecular mass complex in Arabidopsis and in yeast cells, and it interacts in vitro with the yeast Rrp44p and Rrp4p exosome subunits, suggesting that it can participate in evolutionarily conserved interactions that could be essential for the integrity of the exosome complex.
Department of Biological Sciences and the Center for Molecular Genetics, State University of New York at Albany, Albany, New York 12222, USA.
J A
JA
Chekanova
R J
RJ
Shaw
M A
MA
Wills
D A
DA
Belostotsky
D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins; D005095:Exoribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D063326:Exosome Multienzyme Ribonuclease Complex; D015723:Gene Library; D005816:Genetic Complementation Test; D006801:Humans; D016366:Open Reading Frames; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D021122:Protein Subunits; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism*; D012340:RNA, Ribosomal, 5.8S / Q000378:metabolism*; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins
D029681:Arabidopsis Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; D012340:RNA, Ribosomal, 5.8S; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C111978:SKI6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C415835:EXOSC4 protein, human; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; C485933:RRP41 protein, Arabidopsis