Archaeal guide RNAs function in rRNA modification in the eukaryotic nucleus.
In eukaryotes, many Box C/D small nucleolar RNAs base pair with ribosomal RNA through short complementary guide sequences, thereby marking up to 100 individual nucleotides of ribosomal RNA for 2'-O-methylation. Function of the eukaryotic Box C/D RNAs depends upon interaction with at least six known proteins. Box C/D RNAs are not known to exist in Bacteria but were recently identified in Archaea by biochemical analysis and computational genomic screens and have likely evolved independently in Archaea and Eukarya for more than 2000 million years. We have microinjected Box C/D RNAs from Pyrococcus furiosus, a hyperthermophilic archaeon, into the nuclei of oocytes from the aquatic frog Xenopus laevis. Our results show that Box C/D RNAs derived from this prokaryote are retained in the nucleus, localize to nucleoli, and interact with the X. laevis Box C/D RNA binding proteins fibrillarin, Nop56, and Nop58. Furthermore, we have demonstrated the ability of archaeal Box C/D RNAs to direct site-specific 2'-O-methylation of ribosomal RNA. Our studies have revealed the remarkable ability of archaeal Box C/D RNAs to assemble into functional RNA-protein complexes in the eukaryotic nucleus.
Speckmann
Wayne A
WA
Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA.
Li
Zhu-Hong
ZH
Lowe
Todd M
TM
Eddy
Sean R
SR
Terns
Rebecca M
RM
Terns
Michael P
MP
D021581:Active Transport, Cell Nucleus; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000166:cytology / Q000378:metabolism*; D008745:Methylation; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020027:Pyrococcus furiosus / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D019642:RNA, Archaeal / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism*; D029867:Xenopus Proteins / Q000378:metabolism; D014982:Xenopus laevis; D058727:RNA, Small Untranslated
D019642:RNA, Archaeal; D012335:RNA, Ribosomal; D029867:Xenopus Proteins; D058727:RNA, Small Untranslated