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Vol. 10, Issue 7, 2131-2147, July 1999
and
Departments of Small nucleolar RNAs (snoRNAs) are a large family of eukaryotic
RNAs that function within the nucleolus in the biogenesis of ribosomes.
One major class of snoRNAs is the box C/D snoRNAs named for
their conserved box C and box D sequence elements. We have investigated
the involvement of cis-acting sequences and intranuclear
structures in the localization of box C/D snoRNAs to the nucleolus by
assaying the intranuclear distribution of fluorescently labeled U3, U8,
and U14 snoRNAs injected into Xenopus oocyte nuclei.
Analysis of an extensive panel of U3 RNA variants showed that the box
C/D motif, comprised of box C', box D, and the 3' terminal stem of U3,
is necessary and sufficient for the nucleolar localization of U3
snoRNA. Disruption of the elements of the box C/D motif of U8 and U14
snoRNAs also prevented nucleolar localization, indicating that all box
C/D snoRNAs use a common nucleolar-targeting mechanism. Finally, we
found that wild-type box C/D snoRNAs transiently associate with coiled
bodies before they localize to nucleoli and that variant RNAs that lack
an intact box C/D motif are detained within coiled bodies. These
results suggest that coiled bodies play a role in the biogenesis and/or intranuclear transport of box C/D snoRNAs.
Biochemistry and Molecular Biology and
*Genetics, University of Georgia, Athens, Georgia 30602
Corresponding author. E-mail
address: mterns{at}bchiris.bmb.uga.edu.
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