Several snoRNAs are essential for the sequence of
cleavage events required to produce the mature forms
of 18S, 5.8S, and 28S rRNA from the large precursor molecule.
In the absence of U22, mature 18S rRNA fails to accumulate;
U8 snoRNA is essential for accumulation of both 5.8S and
28S rRNA. The mechanisms by which snoRNAs facilitate these
cleavage events is not known and might include direct cleavage
or assisting the rate or efficiency of ribosome assembly.
To learn more about the mechanisms of snoRNA-mediated pre-rRNA
processing, an examination of the kinetics of pre-rRNA
processing in Xenopus oocytes was undertaken.
Correct pre-rRNA processing can be restored in snoRNA-depleted
oocytes following cytoplasmic injection of the corresponding
in vitro-synthesized snoRNA. Analysis of the kinetics of
pre-rRNA processing in these snoRNA-rescue experiments
demonstrated that the rate of accumulation of mature rRNAs
was slower than that seen in untreated oocytes. The snoRNAs
were imported into the nucleus at a rate and overall efficiency
less than that of U1 snRNA, used as a control for import.
However, sufficient levels of snoRNA were present in the
nucleus to yield a functional phenotype (rescue of rRNA
processing) several hours before the snoRNAs were directly
detectable in the nucleus via autoradiography. This indicated
that very low amounts of the snoRNA in the nucleus were
sufficient for rescue. Finally, transcriptional inhibitors
were used to separate transcription and processing. Failure
to rescue snoRNA-mediated processing of pre-accumulated
precursors is consistent with a scenario in which U8 and
U22 must be present during transcription of pre-rRNA.