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The U14 snoRNA is required for 2′-O-methylation of the pre-18S rRNA in Xenopus oocytes

Published online by Cambridge University Press:  01 February 1998

D.A. DUNBAR
Affiliation:
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06520-8040, USA
S.J. BASERGA
Affiliation:
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06520-8040, USA Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520-8040, USA
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Abstract

We have studied the role of the U14 small nucleolar RNA (snoRNA) in pre-rRNA methylation and processing in Xenopus oocytes. Depletion of U14 in Xenopus oocytes was achieved by co-injecting two nonoverlapping antisense oligonucleotides. Focusing on the earliest precursor, depletion experiments revealed that the U14 snoRNA is essential for 2′-O-ribose methylation at nt 427 of the 18S rRNA. Injection of U14-depleted oocytes with specific U14 mutant snoRNAs indicated that conserved domain B, but not domain A, of U14 is required for the methylation reaction. When the effect of U14 on pre-rRNA processing is assayed, we find only modest effects on 18S rRNA levels, and no effect on the type or accumulation of 18S precursors, suggesting a role for U14 in a step in ribosome biogenesis other than cleavage of the pre-rRNA. Xenopus U14 is, therefore, a Box C/D fibrillarin-associated snoRNA that is required for site-specific 2′-O-ribose methylation of pre-rRNA.

Type
Research Article
Information
RNA , Volume 4 , Issue 2 , February 1998 , pp. 195 - 204
Copyright
© 1998 RNA Society

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