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Vol. 11, Issue 9, 2987-2998, September 2000
Department of Cell Biology, University of Massachusetts Medical
School, Worcester, Massachusetts 01655
The Cajal (coiled) body (CB) is a structure enriched in proteins
involved in mRNA, rRNA, and snRNA metabolism. CBs have been shown to
interact with specific histone and snRNA gene loci. To examine the
potential role of CBs in U2 snRNA metabolism, we used a variety of
genomic and oligonucleotide probes to visualize in situ newly
synthesized U2 snRNA relative to U2 loci and CBs. Results demonstrate
that long spacer sequences between U2 coding repeats are transcribed,
supporting other recent evidence that U2 transcription proceeds past
the 3' box. The presence of bright foci of this U2 locus RNA differed
between alleles within the same nucleus; however, this did not
correlate with the loci's association with a CB. Experiments with
specific oligonucleotide probes revealed signal for preU2 RNA within
CBs. PreU2 was also detected in the locus-associated RNA foci, whereas
sequences 3' of preU2 were found only in these foci, not in CBs. This
suggests that a longer primary transcript is processed before entry
into CBs. Although this work shows that direct contact of a U2 locus
with a CB is not simply correlated with RNA at that locus, it provides
the first evidence of new preU2 transcripts within CBs. We also show that, in contrast to CBs, SMN gems do not associate with U2 gene loci
and do not contain preU2. Because other evidence indicates that preU2
is processed in the cytoplasm before assembly into snRNPs,
results point to an involvement of CBs in modification or transport of
preU2 RNA.
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