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Department of Biochemistry, University of Mississippi Medical
Center, Jackson, Mississippi 39216-4505
Previous studies showed that components implicated in pre-rRNA
processing, including U3 small nucleolar (sno)RNA, fibrillarin, nucleolin, and proteins B23 and p52, accumulate in perichromosomal regions and in numerous mitotic cytoplasmic particles, termed nucleolus-derived foci (NDF) between early anaphase and late telophase. The latter structures were analyzed for the presence of pre-rRNA by
fluorescence in situ hybridization using probes for segments of
pre-rRNA with known half-lives. The NDF did not contain the short-lived
5'-external transcribed spacer (ETS) leader segment upstream from the
primary processing site in 47S pre-rRNA. However, the NDF contained
sequences from the 5'-ETS core, 18S, internal transcribed spacer 1 (ITS1), and 28S segments and also had detectable, but significantly
reduced, levels of the 3'-ETS sequence. Northern analyses showed that
in mitotic cells, the latter sequences were present predominantly in
45S-46S pre-rRNAs, indicating that high-molecular weight processing
intermediates are preserved during mitosis. Two additional essential
processing components were also found in the NDF: U8 snoRNA and hPop1
(a protein component of RNase MRP and RNase P). Thus, the NDF
appear to be large complexes containing partially processed pre-rRNA
associated with processing components in which processing has been
significantly suppressed. The NDF may facilitate coordinated assembly
of postmitotic nucleoli.
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