© The Rockefeller University Press,
0021-9525/1998//899 $5.00
The Journal of Cell Biology, Volume 142, Number 4,
, 1998 899-912
Microinjection of Anti-coilin Antibodies Affects the Structure of Coiled Bodies
Fátima Almeida*,
Rainer Saffrich
,
Wilhelm Ansorge
, and
Maria Carmo-Fonseca*
* Institute of Histology and Embryology, Faculty of Medicine, University of Lisbon, 1699 Lisboa Codex, Portugal; and
European Molecular Biology Laboratory, D-69117 Heidelberg, Germany
The coiled body is a distinct subnuclear domain enriched in small nuclear ribonucleoprotein particles (snRNPs) involved in processing of pre-mRNA. Although the function of the coiled body is still unknown, current models propose that it may have a role in snRNP biogenesis, transport, or recycling. Here we describe that anti-coilin antibodies promote a specific disappearance of the coiled body in living human cells, thus providing a novel tool for the functional analysis of this structure. Monoclonal antibodies (mAbs) were raised against recombinant human coilin, the major structural protein of the coiled body. Four mAbs are shown to induce a progressive disappearance of coiled bodies within
6 h after microinjection into the nucleus of HeLa cells. After their disappearance, coiled bodies are not seen to re-form, although injected cells remain viable for at least 3 d. Epitope mapping reveals that the mAbs recognize distinct amino acid motifs scattered along the complete coilin sequence. By 24 and 48 h after injection of antibodies that promote coiled body disappearance, splicing snRNPs are normally distributed in the nucleoplasm, the nucleolus remains unaffected, and the cell cycle progresses normally. Furthermore, cells devoid of coiled bodies for
24 h maintain the ability to splice both adenoviral pre-mRNAs and transiently overexpressed human β-globin transcripts. In conclusion, within the time range of this study, no major nuclear abnormalities are detected after coiled body disappearance.
Key Words: coiled body p80–coilin splicing spliceosomal snRNPs nucleolus
Abbreviations used in this paper: Ad2, adenovirus type 2; BrdU, bromodeoxyuridine; BrUTP, 5-bromo-2'-uridine-5'-triphosphate; Cy5, indodicarbocyanine; MLP, major late promotor; NLS, nuclear localization signal; snoRNA, small nucleolar RNA; snRNP, small nuclear ribonucleoprotein.
Address all correspondence to M. Carmo-Fonseca, Institute of Histology and Embryology, Faculty of Medicine, Av. Prof. Egas Moniz, 1699 Lisboa Codex, Portugal. Tel.: (351) 1 7934340. Fax: (351) 1 7951780. E-mail: hcarmo{at}fm.ul.pt

CiteULike
Complore
Connotea
Del.icio.us
Digg
Facebook
Reddit
Technorati
Twitter What's this?
This article has been cited by other articles:
-
Huranova, M., Hnilicova, J., Fleischer, B., Cvackova, Z., Stanek, D.
(2009). A mutation linked to retinitis pigmentosa in HPRP31 causes protein instability and impairs its interactions with spliceosomal snRNPs. Hum Mol Genet
18: 2014-2023
[Abstract]
[Full Text]
-
Guffanti, E., Kittur, N., Brodt, Z. N., Polotsky, A. J., Kuokkanen, S. M., Heller, D. S., Young, S. L., Santoro, N., Meier, U. T.
(2008). Nuclear pore complex proteins mark the implantation window in human endometrium. J. Cell Sci.
121: 2037-2045
[Abstract]
[Full Text]
-
Stanek, D., Pridalova-Hnilicova, J., Novotny, I., Huranova, M., Blazikova, M., Wen, X., Sapra, A. K., Neugebauer, K. M.
(2008). Spliceosomal Small Nuclear Ribonucleoprotein Particles Repeatedly Cycle through Cajal Bodies. Mol. Biol. Cell
19: 2534-2543
[Abstract]
[Full Text]
-
Rino, J., Desterro, J. M. P., Pacheco, T. R., Gadella, T. W. J. Jr., Carmo-Fonseca, M.
(2008). Splicing Factors SF1 and U2AF Associate in Extraspliceosomal Complexes. Mol. Cell. Biol.
28: 3045-3057
[Abstract]
[Full Text]
-
Petri, S., Grimmler, M., Over, S., Fischer, U., Gruss, O. J.
(2007). Dephosphorylation of survival motor neurons (SMN) by PPM1G/PP2C{gamma} governs Cajal body localization and stability of the SMN complex. JCB
179: 451-465
[Abstract]
[Full Text]
-
Cardinale, S., Cisterna, B., Bonetti, P., Aringhieri, C., Biggiogera, M., Barabino, S. M.L.
(2007). Subnuclear Localization and Dynamics of the Pre-mRNA 3' End Processing Factor Mammalian Cleavage Factor I 68-kDa Subunit. Mol. Biol. Cell
18: 1282-1292
[Abstract]
[Full Text]
-
Kittur, N., Darzacq, X., Roy, S., Singer, R. H., Meier, U. T.
(2006). Dynamic association and localization of human H/ACA RNP proteins. RNA
12: 2057-2062
[Abstract]
[Full Text]
-
Klingauf, M., Stanek, D., Neugebauer, K. M.
(2006). Enhancement of U4/U6 Small Nuclear Ribonucleoprotein Particle Association in Cajal Bodies Predicted by Mathematical Modeling. Mol. Biol. Cell
17: 4972-4981
[Abstract]
[Full Text]
-
Lemm, I., Girard, C., Kuhn, A. N., Watkins, N. J., Schneider, M., Bordonne, R., Luhrmann, R.
(2006). Ongoing U snRNP Biogenesis Is Required for the Integrity of Cajal Bodies. Mol. Biol. Cell
17: 3221-3231
[Abstract]
[Full Text]
-
Darzacq, X., Kittur, N., Roy, S., Shav-Tal, Y., Singer, R. H., Meier, U. T.
(2006). Stepwise RNP assembly at the site of H/ACA RNA transcription in human cells. JCB
173: 207-218
[Abstract]
[Full Text]
-
Sun, J., Xu, H., Subramony, S. H., Hebert, M. D.
(2005). Interactions between Coilin and PIASy partially link Cajal bodies to PML bodies. J. Cell Sci.
118: 4995-5003
[Abstract]
[Full Text]
-
Feng, W., Gubitz, A. K., Wan, L., Battle, D. J., Dostie, J., Golembe, T. J., Dreyfuss, G.
(2005). Gemins modulate the expression and activity of the SMN complex. Hum Mol Genet
14: 1605-1611
[Abstract]
[Full Text]
-
TAVANEZ, J. P., CALADO, P., BRAGA, J., LAFARGA, M., CARMO-FONSECA, M.
(2005). In vivo aggregation properties of the nuclear poly(A)-binding protein PABPN1. RNA
11: 752-762
[Abstract]
[Full Text]
-
Tanackovic, G., Kramer, A.
(2005). Human Splicing Factor SF3a, but Not SF1, Is Essential for Pre-mRNA Splicing In Vivo. Mol. Biol. Cell
16: 1366-1377
[Abstract]
[Full Text]
-
Braga, J., Desterro, J. M.P., Carmo-Fonseca, M.
(2004). Intracellular Macromolecular Mobility Measured by Fluorescence Recovery after Photobleaching with Confocal Laser Scanning Microscopes. Mol. Biol. Cell
15: 4749-4760
[Abstract]
[Full Text]
-
Stanek, D., Neugebauer, K. M.
(2004). Detection of snRNP assembly intermediates in Cajal bodies by fluorescence resonance energy transfer. JCB
166: 1015-1025
[Abstract]
[Full Text]
-
Nesic, D., Tanackovic, G., Kramer, A.
(2004). A role for Cajal bodies in the final steps of U2 snRNP biogenesis. J. Cell Sci.
117: 4423-4433
[Abstract]
[Full Text]
-
Berciano, M. T., Villagra, N. T., Ojeda, J. L., Navascues, J., Gomes, A., Lafarga, M., Carmo-Fonseca, M.
(2004). Oculopharyngeal muscular dystrophy-like nuclear inclusions are present in normal magnocellular neurosecretory neurons of the hypothalamus. Hum Mol Genet
13: 829-838
[Abstract]
[Full Text]
-
Zhu, Y., Tomlinson, R. L., Lukowiak, A. A., Terns, R. M., Terns, M. P.
(2004). Telomerase RNA Accumulates in Cajal Bodies in Human Cancer Cells. Mol. Biol. Cell
15: 81-90
[Abstract]
[Full Text]
-
Polak, P. E., Simone, F., Kaberlein, J. J., Luo, R. T., Thirman, M. J.
(2003). ELL and EAF1 are Cajal Body Components That Are Disrupted in MLL-ELL Leukemia. Mol. Biol. Cell
14: 1517-1528
[Abstract]
[Full Text]
-
Stanek, D., Rader, S. D., Klingauf, M., Neugebauer, K. M.
(2003). Targeting of U4/U6 small nuclear RNP assembly factor SART3/p110 to Cajal bodies. JCB
160: 505-516
[Abstract]
[Full Text]
-
Boisvert, F.-M., Cote, J., Boulanger, M.-C., Cleroux, P., Bachand, F., Autexier, C., Richard, S.
(2002). Symmetrical dimethylarginine methylation is required for the localization of SMN in Cajal bodies and pre-mRNA splicing. JCB
159: 957-969
[Abstract]
[Full Text]
-
Calapez, A., Pereira, H. M., Calado, A., Braga, J., Rino, J., Carvalho, C., Tavanez, J. P., Wahle, E., Rosa, A. C., Carmo-Fonseca, M.
(2002). The intranuclear mobility of messenger RNA binding proteins is ATP dependent and temperature sensitive. JCB
159: 795-805
[Abstract]
[Full Text]
-
Roberts, S., Hillman, M. L., Knight, G. L., Gallimore, P. H.
(2002). The ND10 Component Promyelocytic Leukemia Protein Relocates to Human Papillomavirus Type 1 E4 Intranuclear Inclusion Bodies in Cultured Keratinocytes and in Warts. J. Virol.
77: 673-684
[Abstract]
[Full Text]
-
Gatfield, D., Izaurralde, E.
(2002). REF1/Aly and the additional exon junction complex proteins are dispensable for nuclear mRNA export. JCB
159: 579-588
[Abstract]
[Full Text]
-
Chatterjee, T. K., Fisher, R. A.
(2002). RGS12TS-S Localizes at Nuclear Matrix-Associated Subnuclear Structures and Represses Transcription: Structural Requirements for Subnuclear Targeting and Transcriptional Repression. Mol. Cell. Biol.
22: 4334-4345
[Abstract]
[Full Text]
-
Shiels, C., Islam, S. A., Vatcheva, R., Sasieni, P., Sternberg, M. J. E., Freemont, P. S., Sheer, D.
(2002). PML bodies associate specifically with the MHC gene cluster in interphase nuclei. J. Cell Sci.
114: 3705-3716
[Abstract]
[Full Text]
-
Tucker, K. E., Berciano, M. T., Jacobs, E. Y., LePage, D. F., Shpargel, K. B., Rossire, J. J., Chan, E. K.L., Lafarga, M., Conlon, R. A., Matera, A. G.
(2001). Residual Cajal bodies in coilin knockout mice fail to recruit Sm snRNPs and SMN, the spinal muscular atrophy gene product. JCB
154: 293-308
[Abstract]
[Full Text]
-
Shopland, L. S., Byron, M., Stein, J. L., Lian, J. B., Stein, G. S., Lawrence, J. B.
(2001). Replication-dependent Histone Gene Expression Is Related to Cajal Body (CB) Association but Does Not Require Sustained CB Contact. Mol. Biol. Cell
12: 565-576
[Abstract]
[Full Text]
-
Vautier, D, Chesne, P, Cunha, C, Calado, A, Renard, J., Carmo-Fonseca, M
(2001). Transcription-dependent nucleocytoplasmic distribution of hnRNP A1 protein in early mouse embryos. J. Cell Sci.
114: 1521-1531
[Abstract]
-
Platani, M., Goldberg, I., Swedlow, J. R., Lamond, A. I.
(2000). In Vivo Analysis of Cajal Body Movement, Separation, and Joining in Live Human Cells. JCB
151: 1561-1574
[Abstract]
[Full Text]
-
Herold, A., Suyama, M., Rodrigues, J. P., Braun, I. C., Kutay, U., Carmo-Fonseca, M., Bork, P., Izaurralde, E.
(2000). TAP (NXF1) Belongs to a Multigene Family of Putative RNA Export Factors with a Conserved Modular Architecture. Mol. Cell. Biol.
20: 8996-9008
[Abstract]
[Full Text]
-
Zhao, J., Kennedy, B. K., Lawrence, B. D., Barbie, D. A., Matera, A. G., Fletcher, J. A., Harlow, E.
(2000). NPAT links cyclin E-Cdk2 to the regulation of replication-dependent histone gene transcription. Genes Dev.
14: 2283-2297
[Abstract]
[Full Text]
-
Ma, T., Van Tine, B. A., Wei, Y., Garrett, M. D., Nelson, D., Adams, P. D., Wang, J., Qin, J., Chow, L. T., Harper, J. W.
(2000). Cell cycle-regulated phosphorylation of p220NPAT by cyclin E/Cdk2 in Cajal bodies promotes histone gene transcription. Genes Dev.
14: 2298-2313
[Abstract]
[Full Text]
-
Nayler, O., Hartmann, A. M., Stamm, S.
(2000). The ER Repeat Protein Yt521-B Localizes to a Novel Subnuclear Compartment. JCB
150: 949-962
[Abstract]
[Full Text]
-
Isaac, C., Marsh, K. L., Paznekas, W. A., Dixon, J., Dixon, M. J., Jabs, E. W., Meier, U. T.
(2000). Characterization of the Nucleolar Gene Product, Treacle, in Treacher Collins Syndrome. Mol. Biol. Cell
11: 3061-3071
[Abstract]
[Full Text]
-
Lewis, J. D., Tollervey, D.
(2000). Like Attracts Like: Getting RNA Processing Together in the Nucleus. Science
288: 1385-1389
[Abstract]
[Full Text]
-
Mermoud, J. E., Costanzi, C., Pehrson, J. R., Brockdorff, N.
(1999). Histone Macroh2a1.2 Relocates to the Inactive X Chromosome after Initiation and Propagation of X-Inactivation. JCB
147: 1399-1408
[Abstract]
[Full Text]
-
Carvalho, T., Almeida, F., Calapez, A., Lafarga, M., Berciano, M. T., Carmo-Fonseca, M.
(1999). The Spinal Muscular Atrophy Disease Gene Product, Smn: A Link between Snrnp Biogenesis and the Cajal (Coiled) Body. JCB
147: 715-728
[Abstract]
[Full Text]
-
Bellini, M., Gall, J. G.
(1999). Coilin Shuttles between the Nucleus and Cytoplasm In Xenopus Oocytes. Mol. Biol. Cell
10: 3425-3434
[Abstract]
[Full Text]
-
Boudonck, K., Dolan, L., Shaw, P. J.
(1999). The Movement of Coiled Bodies Visualized in Living Plant Cells by the Green Fluorescent Protein. Mol. Biol. Cell
10: 2297-2307
[Abstract]
[Full Text]