Human RBM28 protein is a specific nucleolar component of the spliceosomal snRNPs (original) (raw)

Abstract

The biogenesis of spliceosomal small nuclear RNAs (snRNAs) involves organized translocations between the cytoplasm and certain nuclear domains, such as Cajal bodies and nucleoli. Here we identify human RBM28 protein as a novel snRNP component, based on affinity selection of U6 small nuclear ribonucleoprotein (snRNP). As shown by immunofluorescence, RBM28 is a nucleolar protein. Anti-RBM28 immunoprecipitation from HeLa cell lysates revealed that this protein specifically associates with U1, U2, U4, U5, and U6 snRNAs. Our data provide the first evidence that RBM28 is a common nucleolar component of the spliceosomal ribonucleoprotein complexes, possibly coordinating their transition through the nucleolus.

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Corresponding author albrecht.bindereif@chemie.bio.uni-giessen.de


References

Achsel, T., Brahms, H., Kastner, B., Bachi, A., Wilm, M., and Lührmann, R. (1999). A doughnut-shaped heteromer of human Sm-like proteins binds to the 3′-end of U6 snRNA, thereby facilitating U4/U6 duplex formation in vitro. EMBO J.18, 5789–5802.10.1093/emboj/18.20.5789Search in Google Scholar PubMed PubMed Central

Bell, M., Schreiner, S., Damianov, A., Reddy, R., and Bindereif, A. (2002). p110, a novel human U6 snRNP protein and U4/U6 snRNP recycling factor. EMBO J.21, 2724–2735.10.1093/emboj/21.11.2724Search in Google Scholar PubMed PubMed Central

Berges, T., Petfalski, E., Tollervey, D., and Hurt, E.C. (1994). Synthetic lethality with fibrillarin identifies NOP77p, a nucleolar protein required for pre-rRNA processing and modification. EMBO J.13, 3136–3148.10.1002/j.1460-2075.1994.tb06612.xSearch in Google Scholar PubMed PubMed Central

Booth, B.L. Jr. and Pugh, B.F. (1997). Identification and characterization of a nuclease specific for the 3′ end of the U6 small nuclear RNA. J. Biol. Chem.272, 984–991.10.1074/jbc.272.2.984Search in Google Scholar PubMed

Damianov, A., Schreiner, S., and Bindereif, A. (2004). Recycling of the U12-type spliceosome requires p110, a component of the U6atac snRNP. Mol. Cell. Biol.24, 1700–1708.10.1128/MCB.24.4.1700-1708.2004Search in Google Scholar PubMed PubMed Central

Darzacq, X., Jady, B.E., Verheggen, C., Kiss, A.M., Bertrand, E., and Kiss, T. (2002). Cajal body-specific small nuclear RNAs: a novel class of 2′-O-methylation and pseudouridylation guide RNAs. EMBO J.21, 2746–2756.10.1093/emboj/21.11.2746Search in Google Scholar PubMed PubMed Central

Dignam, J.D., Lebovitz, R.M., and Roeder, R.G. (1983). Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res.11, 1475–1489.10.1093/nar/11.5.1475Search in Google Scholar PubMed PubMed Central

Ganot, P., Jady, B.E., Bortolin, M.L., Darzacq, X., and Kiss, T. (1999). Nucleolar factors direct the 2′-_O_-ribose methylation and pseudouridylation of U6 spliceosomal RNA. Mol. Cell. Biol.19, 6906–6917.10.1128/MCB.19.10.6906Search in Google Scholar PubMed PubMed Central

Gerbi, S.A. and Lange, T.S. (2002). All small nuclear RNAs (snRNAs) of the [U4/U6.U5] Tri-snRNP localize to nucleoli; identification of the nucleolar localization element of U6 snRNA. Mol. Biol. Cell13, 3123–3137.Search in Google Scholar

Gerbi, S.A., Borovjagin, A.V., Odreman, F.E., and Lange, T.S. (2003). U4 snRNA nucleolar localization requires the NHPX/15.5-kDa protein binding site but not Sm protein or U6 snRNA association. J. Cell. Biol.162, 821–832.Search in Google Scholar

Jady, B.E. and Kiss, T. (2001). A small nucleolar guide RNA functions both in 2′-_O_-ribose methylation and pseudouridylation of the U5 spliceosomal RNA. EMBO J.20, 541–551.10.1093/emboj/20.3.541Search in Google Scholar PubMed PubMed Central

Jady, B.E., Darzacq, X., Tucker, K.E., Matera, A.G., Bertrand, E., and Kiss, T. (2003). Modification of Sm small nuclear RNAs occurs in the nucleoplasmic Cajal body following import from the cytoplasm. EMBO J.22, 1878–1888.10.1093/emboj/cdg187Search in Google Scholar PubMed PubMed Central

Kiss, T. (2004). Biogenesis of small nuclear RNPs. J. Cell Sci.117, 5949–5951.10.1242/jcs.01487Search in Google Scholar

Kiss, A.M., Jady, B.E., Darzacq, X., Verheggen, C., Bertrand, E., and Kiss, T. (2002). A Cajal body-specific pseudouridylation guide RNA is composed of two box H/ACA snoRNA-like domains. Nucleic Acids Res.30, 4643–4649.10.1093/nar/gkf592Search in Google Scholar

Lund, E. and Dahlberg, J.E. (1992). Cyclic 2′,3′-phosphates and nontemplated nucleotides at the 3′ end of spliceosomal U6 small nuclear RNA's. Science255, 327–330.10.1126/science.1549778Search in Google Scholar

Marchler-Bauer, A. and Bryant, S.H. (2004). CD-Search: protein domain annotations on the fly. Nucleic Acids Res.32, W327–W331.10.1093/nar/gkh454Search in Google Scholar

Mattaj, I.W. (1986). Cap trimethylation of U snRNA is cytoplasmic and dependent on U snRNP protein binding. Cell46, 905–911.10.1016/0092-8674(86)90072-3Search in Google Scholar

Mougin, A., Gottschalk, A., Fabrizio, P., Lührmann, R., and Branlant, C. (2002). Direct probing of RNA structure and RNA-protein interactions in purified HeLa cells and yeast spliceosomal U4/U6.U5 tri-snRNP particles. J. Mol. Biol.317, 631–649.Search in Google Scholar

Palacios, I., Hetzer, M., Adam, S.A., and Mattaj, I.W. (1997). Nuclear import of U snRNPs requires importin β. EMBO J.16, 6783–6792.10.1093/emboj/16.22.6783Search in Google Scholar

Pannone, B.K., Xue, D., and Wolin, S.L. (1998). A role for the yeast La protein in U6 snRNP assembly: evidence that the La protein is a molecular chaperone for RNA polymerase III transcripts. EMBO J.17, 7442–7453.10.1093/emboj/17.24.7442Search in Google Scholar

Seipelt, R.L., Zheng, B., Asuru, A., and Rymond, B.C. (1999). U1 snRNA is cleaved by RNase III and processed through an Sm site-dependent pathway. Nucleic Acids Res.27, 587–595.10.1093/nar/27.2.587Search in Google Scholar

Singh, R. and Reddy, R. (1989). Gamma-monomethyl phosphate: a cap structure in spliceosomal U6 small nuclear RNA. Proc. Natl. Acad. Sci. USA86, 8280–8283.10.1073/pnas.86.21.8280Search in Google Scholar

Sleeman, J.E. and Lamond, A.I. (1999). Newly assembled snRNPs associate with coiled bodies before speckles, suggesting a nuclear snRNP maturation pathway. Curr. Biol.9, 1065–1074.10.1016/S0960-9822(99)80475-8Search in Google Scholar

Sun, C. and Woolford, J.L. Jr. (1994). The yeast NOP4 gene product is an essential nucleolar protein required for pre-rRNA processing and accumulation of 60S ribosomal subunits. EMBO J.13, 3127–3135.10.1002/j.1460-2075.1994.tb06611.xSearch in Google Scholar

Sun, C. and Woolford, J.L. Jr. (1997). The yeast nucleolar protein Nop4p contains four RNA recognition motifs necessary for ribosome biogenesis. J. Biol. Chem.272, 25345–25352.10.1074/jbc.272.40.25345Search in Google Scholar

Terns, M.P., Lund, E., and Dahlberg, J.E. (1992). 3′-End-dependent formation of U6 small nuclear ribonucleoprotein particles in Xenopus laevis oocyte nuclei. Mol. Cell. Biol.12, 3032–3040.Search in Google Scholar

Trippe, R., Sandrock, B., and Benecke, B.J. (1998). A highly specific terminal uridylyl transferase modifies the 3′-end of U6 small nuclear RNA. Nucleic Acids Res.26, 3119–3126.10.1093/nar/26.13.3119Search in Google Scholar

Trippe, R., Guschina, E., Hossbach, M., Urlaub, H., Lührmann, R., and Benecke, B.H. (2006). Identification, cloning, and functional analysis of the human U6 snRNA-specific terminal uridylyl transferase. RNA12, 1494–1504.10.1261/rna.87706Search in Google Scholar

Tycowski, K.T., You, Z.H., Graham, P.J., and Steitz, J.A. (1998). Modification of U6 spliceosomal RNA is guided by other small RNAs. Mol. Cell2, 629–638.10.1016/S1097-2765(00)80161-6Search in Google Scholar

Will, C.L. and Lührmann, R. (2001). Spliceosomal UsnRNP biogenesis, structure and function. Curr. Opin. Cell Biol.13, 290–301.10.1016/S0955-0674(00)00211-8Search in Google Scholar

Wolff, T. and Bindereif, A. (1992). Reconstituted mammalian U4/U6 snRNP complements splicing: a mutational analysis. EMBO J.11, 345–359.10.1002/j.1460-2075.1992.tb05057.xSearch in Google Scholar PubMed PubMed Central

Yong, J., Wan, L., and Dreyfuss, G. (2004). Why do cells need an assembly machine for RNA-protein complexes? Trends Cell Biol.14, 226–232.Search in Google Scholar

Published Online: 2006-11-02

Published in Print: 2006-10-01

©2006 by Walter de Gruyter Berlin New York