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Vol. 11, Issue 7, 2419-2428, July 2000
Division of Biology and Medicine, Brown University, Providence,
Rhode Island 02912
Recent studies on the 2'-O-methylation and pseudouridylation of U6
small nuclear RNA (snRNA) hypothesize that these posttranscriptional modifications might occur in the nucleolus. In this report, we present
direct evidence for the nucleolar localization of U6 snRNA and analyze
the kinetics of U6 nucleolar localization after injection of in vitro
transcribed fluorescein-labeled transcripts into Xenopus laevis oocytes. In contrast to U3 small nucleolar RNA (snoRNA) which developed strong nucleolar labeling over 4 h and maintained strong nucleolar signals through 24 h, U6 snRNA localized to
nucleoli immediately after injection, but nucleolar staining decreased after 4 h. By 24 h after injection of U6 snRNA, only weak
nucleolar signals were observed. Unlike the time-dependent profile of
strong nucleolar localization of U6 snRNA or U3 snoRNA, injection of fluorescein-labeled U2 snRNA gave weak nucleolar staining at all times
throughout a 24-h period; U2 snRNA modifications are believed to occur
outside of the nucleolus. The notion that the decrease of U6 signals
over time was due to its trafficking out of nucleoli and not to
transcript degradation was supported by the demonstration of U6 snRNA
stability over time. Therefore, in contrast to snoRNAs like U3, U6
snRNA transiently passes through nucleoli.
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