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Published online 25 August 2003. doi:10.1083/jcb.200301071
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© The Rockefeller University Press, 0021-9525/2003/9/821 $5.00
The Journal of Cell Biology, Volume 162, Number 5, 821-832

Article

 U4 snRNA nucleolar localization requires the NHPX/15.5-kD protein binding site but not Sm protein or U6 snRNA association

Susan A. Gerbi, Anton V. Borovjagin, Federico E. Odreman and Thilo Sascha Lange

Division of Biology and Medicine, Brown University, Providence, RI 02912

Address correspondence to Thilo Sascha Lange, Division of Biology and Medicine, Brown University, 69 Brown St., J.W. Wilson Laboratory, Providence, RI 02912. Tel.: (401) 863-2420. Fax: (401) 863-1348. email: Thilo_Lange{at}brown.edu

All small nuclear RNAs (snRNAs) of the [U4/U6.U5] tri-snRNP localize transiently to nucleoli, as visualized by microscopy after injection of fluorescein-labeled transcripts into Xenopus laevis oocyte nuclei. Here, we demonstrate that these RNAs traffic to nucleoli independently of one another, because U4 snRNA deleted in the U6 base-pairing region still localizes to nucleoli. Furthermore, depletion of endogenous U6 snRNA does not affect nucleolar localization of injected U4 or U5. The wild-type U4 transcripts used here are functional: they exhibit normal nucleocytoplasmic traffic, associate with Sm proteins, form the [U4/U6] di-snRNP, and localize to nucleoli and Cajal bodies. The nucleolar localization element (NoLE) of U4 snRNA was mapped by mutagenesis. Neither the 5'-cap nor the 3'-region of U4, which includes the Sm protein binding site, are essential for nucleolar localization. The only region in U4 snRNA required for nucleolar localization is the 5'-proximal stem loop, which contains the binding site for the NHPX/15.5-kD protein. Even mutation of just five nucleotides, essential for binding this protein, impaired U4 nucleolar localization. Intriguingly, the NHPX/15.5-kD protein also binds the nucleolar localization element of box C/D small nucleolar RNAs, suggesting that this protein might mediate nucleolar localization of several small RNAs.

Key Words: U4 snRNA; nucleolar localization elements (NoLEs); NHPX/15.5-kD (Snu13) protein; [U4/U6.U5] tri-snRNP; Sm proteins

Abbreviations used in this paper: NoLE, nucleolar localization element; pol, polymerase; scaRNA, small Cajal body–specific RNA. snRNA, small nuclear RNA; snRNP; small nuclear RNP; snoRNA, small nucleolar RNA.


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