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Published online September 10, 2007
doi:10.1083/jcb.200706134
The Journal of Cell Biology, Vol. 178, No. 6, 937-949
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Patel et al.
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Article

 Splicing-independent recruitment of spliceosomal small nuclear RNPs to nascent RNA polymerase II transcripts

Snehal Bhikhu Patel1, Natalya Novikova2, and Michel Bellini2

1 Department of Biochemistry, College of Medicine and 2 Department of Cell and Developmental Biology, School of Molecular and Cellular Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801

Correspondence to Michel Bellini: bellini{at}life.uiuc.edu

In amphibian oocytes, most lateral loops of the lampbrush chromosomes correspond to active transcriptional sites for RNA polymerase II. We show that newly assembled small nuclear ribonucleoprotein (RNP [snRNP]) particles, which are formed upon cytoplasmic injection of fluorescently labeled spliceosomal small nuclear RNAs (snRNAs), target the nascent transcripts of the chromosomal loops. With this new targeting assay, we demonstrate that nonfunctional forms of U1 and U2 snRNAs still associate with the active transcriptional units. In particular, we find that their association with nascent RNP fibrils is independent of their base pairing with pre–messenger RNAs. Additionally, stem loop I of the U1 snRNA is identified as a discrete domain that is both necessary and sufficient for association with nascent transcripts. Finally, in oocytes deficient in splicing, the recruitment of U1, U4, and U5 snRNPs to transcriptional units is not affected. Collectively, these data indicate that the recruitment of snRNPs to nascent transcripts and the assembly of the spliceosome are uncoupled events.

Abbreviations used in this paper: BPS, branch point sequence; CB, Cajal body; DIC, differential interference contrast; EJC, exon junction complex; IGC, interchromatin granule cluster; LBC, lampbrush chromosome; RNAP, RNA polymerase; snRNA, small nuclear RNA; snRNP, small nuclear RNP; SS, splice site.


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