© The Rockefeller University Press,
0021-9525/1997//965 $5.00
The Journal of Cell Biology, Volume 137, Number 5,
, 1997 965-974
The Dynamic Organization of the Perinucleolar Compartment in the Cell Nucleus
Sui Huang*,
Thomas J. Deerinck
,
Mark H. Ellisman
, and
David L. Spector*
* Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724; and
The National Center for Microscopy and Imaging Research at San Diego, University of California, La Jolla, California 92093
The perinucleolar compartment (PNC) is a unique nuclear structure preferentially localized at the periphery of the nucleolus. Several small RNAs transcribed by RNA polymerase III (e.g., the Y RNAs, MRP RNA, and RNase P H1 RNA) and the polypyrimidine tract binding protein (PTB; hnRNP I) have thus far been identified in the PNC (Ghetti, A., S. PinolRoma, W.M. Michael, C. Morandi, and G. Dreyfuss. 1992. Nucleic Acids Res. 20:3671–3678; Matera, A.G., M.R. Frey, K. Margelot, and S.L. Wolin. 1995. J. Cell Biol. 129:1181–1193; Lee, B., A.G. Matera, D.C. Ward, and J. Craft. 1996. Proc. Natl. Acad. Sci. USA. 93: 11471–11476). In this report, we have further characterized this structure in both fixed and living cells. Detection of the PNC in a large number of human cancer and normal cells showed that PNCs are much more prevalent in cancer cells. Analysis through the cell cycle using immunolabeling with a monoclonal antibody, SH54, specifically recognizing PTB, demonstrated that the PNC dissociates at the beginning of mitosis and reforms at late telophase in the daughter nuclei. To visualize the PNC in living cells, a fusion protein between PTB and green fluorescent protein (GFP) was generated. Time lapse studies revealed that the size and shape of the PNC is dynamic over time. In addition, electron microscopic examination in optimally fixed cells revealed that the PNC is composed of multiple strands, each measuring
80–180 nm diam. Some of the strands are in direct contact with the surface of the nucleolus. Furthermore, analysis of the sequence requirement for targeting PTB to the PNC using a series of deletion mutants of the GFP–PTB fusion protein showed that at least three RRMs at either the COOH or NH2 terminus are required for the fusion protein to be targeted to the PNC. This finding suggests that RNA binding may be necessary for PTB to be localized in the PNC.
1. Abbreviations used in this paper: GFP, green fluorescent protein; PML, promyelocyte; PNC, perinucleolar complex; POD, PML oncogenic domain; PTB, polypyrimidine tract binding protein; RRM, RNA recognition motif.
We would like to express our special gratitude to Tamara Howard for her excellent technical assistance. We would also like to thank Dr. G. Dreyfuss (University of Pennsylvania, Philadelphia, PA) and his colleagues for confirming the specificity of the antibody SH54 to PTB, and Dr. E. Wimmer (S.U.N.Y., Stony Brook, NY) for the gift of the human PTB cDNA clone. We are very grateful to Drs. D. Helfman (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY), G. Matera (Case Western Reserve University, Cleveland, OH), P. Mintz (Cold Spring Harbor Laboratory), and T. Misteli (Cold Spring Harbor Laboratory) for their helpful comments on the manuscript.
D.L. Spector is supported by a grant from the National Institutes of Health (GM42694).
Please address all correspondence to Dr. Sui Huang, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724. Tel.: (516) 367-8478; Fax: (516) 367-8876; E-mail: huang{at}cshl.org

CiteULike
Complore
Connotea
Del.icio.us
Digg
Facebook
Reddit
Technorati
Twitter What's this?
This article has been cited by other articles:
-
Norton, J. T., Titus, S. A., Dexter, D., Austin, C. P., Zheng, W., Huang, S.
(2009). Automated High-Content Screening for Compounds That Disassemble the Perinucleolar Compartment. J Biomol Screen
14: 1045-1053
[Abstract]
-
Norton, J. T., Wang, C., Gjidoda, A., Henry, R. W., Huang, S.
(2009). The Perinucleolar Compartment Is Directly Associated with DNA. J. Biol. Chem.
284: 4090-4101
[Abstract]
[Full Text]
-
Ernoult-Lange, M., Wilczynska, A., Harper, M., Aigueperse, C., Dautry, F., Kress, M., Weil, D.
(2009). Nucleocytoplasmic Traffic of CPEB1 and Accumulation in Crm1 Nucleolar Bodies. Mol. Biol. Cell
20: 176-187
[Abstract]
[Full Text]
-
Wang, C., Norton, J. T., Ghosh, S., Kim, J., Fushimi, K., Wu, J. Y., Stack, M. S., Huang, S.
(2008). Polypyrimidine Tract-binding Protein (PTB) Differentially Affects Malignancy in a Cell Line-dependent Manner. J. Biol. Chem.
283: 20277-20287
[Abstract]
[Full Text]
-
Florez, P. M., Sessions, O. M., Wagner, E. J., Gromeier, M., Garcia-Blanco, M. A.
(2005). The Polypyrimidine Tract Binding Protein Is Required for Efficient Picornavirus Gene Expression and Propagation. J. Virol.
79: 6172-6179
[Abstract]
[Full Text]
-
Shav-Tal, Y., Blechman, J., Darzacq, X., Montagna, C., Dye, B. T., Patton, J. G., Singer, R. H., Zipori, D.
(2005). Dynamic Sorting of Nuclear Components into Distinct Nucleolar Caps during Transcriptional Inhibition. Mol. Biol. Cell
16: 2395-2413
[Abstract]
[Full Text]
-
Kamath, R. V., Thor, A. D., Wang, C., Edgerton, S. M., Slusarczyk, A., Leary, D.J., Wang, J., Wiley, E.L., Jovanovic, B., Wu, Q., Nayar, R., Kovarik, P., Shi, F., Huang, S.
(2005). Perinucleolar Compartment Prevalence Has an Independent Prognostic Value for Breast Cancer. Cancer Res.
65: 246-253
[Abstract]
[Full Text]
-
Eilbracht, J., Reichenzeller, M., Hergt, M., Schnolzer, M., Heid, H., Stohr, M., Franke, W. W., Schmidt-Zachmann, M. S.
(2004). NO66, a Highly Conserved Dual Location Protein in the Nucleolus and in a Special Type of Synchronously Replicating Chromatin. Mol. Biol. Cell
15: 1816-1832
[Abstract]
[Full Text]
-
Hall, M. P., Huang, S., Black, D. L.
(2004). Differentiation-induced Colocalization of the KH-type Splicing Regulatory Protein with Polypyrimidine Tract Binding Protein and the c-src Pre-mRNA. Mol. Biol. Cell
15: 774-786
[Abstract]
[Full Text]
-
Wang, C., Politz, J. C., Pederson, T., Huang, S.
(2003). RNA Polymerase III Transcripts and the PTB Protein Are Essential for the Integrity of the Perinucleolar Compartment. Mol. Biol. Cell
14: 2425-2435
[Abstract]
[Full Text]
-
Back, S. H., Shin, S., Jang, S. K.
(2002). Polypyrimidine Tract-binding Proteins Are Cleaved by Caspase-3 during Apoptosis. J. Biol. Chem.
277: 27200-27209
[Abstract]
[Full Text]
-
Ko, T. K., Kelly, E., Pines, J.
(2002). CrkRS: a novel conserved Cdc2-related protein kinase that colocalises with SC35 speckles. J. Cell Sci.
114: 2591-2603
[Abstract]
[Full Text]
-
Back, S. H., Kim, Y. K., Kim, W. J., Cho, S., Oh, H. R., Kim, J.-E., Jang, S. K.
(2002). Translation of Polioviral mRNA Is Inhibited by Cleavage of Polypyrimidine Tract-Binding Proteins Executed by Polioviral 3Cpro. J. Virol.
76: 2529-2542
[Abstract]
[Full Text]
-
Yuan, X., Davydova, N., Conte, M. R., Curry, S., Matthews, S.
(2002). Chemical shift mapping of RNA interactions with the polypyrimidine tract binding protein. Nucleic Acids Res
30: 456-462
[Abstract]
[Full Text]
-
Zang, W.-Q., Li, B., Huang, P.-Y., Lai, M. M. C., Yen, T. S. B.
(2001). Role of Polypyrimidine Tract Binding Protein in the Function of the Hepatitis B Virus Posttranscriptional Regulatory Element. J. Virol.
75: 10779-10786
[Abstract]
[Full Text]
-
Misteli, T
(2000). Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function. J. Cell Sci.
113: 1841-1849
[Abstract]
-
Chen, T., Boisvert, F.-M., Bazett-Jones, D. P., Richard, S.
(1999). A Role for the GSG Domain in Localizing Sam68 to Novel Nuclear Structures in Cancer Cell Lines. Mol. Biol. Cell
10: 3015-3033
[Abstract]
[Full Text]
-
Huang, S., Deerinck, T. J., Ellisman, M. H., Spector, D. L.
(1998). The Perinucleolar Compartment and Transcription. JCB
143: 35-47
[Abstract]
[Full Text]
-
Saurin, A. J., Shiels, C., Williamson, J., Satijn, D. P.E., Otte, A. P., Sheer, D., Freemont, P. S.
(1998). The Human Polycomb Group Complex Associates with Pericentromeric Heterochromatin to Form a Novel Nuclear Domain. JCB
142: 887-898
[Abstract]
[Full Text]
-
Lamond, A. I., Earnshaw, W. C.
(1998). Structure and Function in the Nucleus. Science
280: 547-553
[Abstract]
[Full Text]
-
Fabini, G., Raijmakers, R., Hayer, S., Fouraux, M. A., Pruijn, G. J. M., Steiner, G.
(2001). The Heterogeneous Nuclear Ribonucleoproteins I and K Interact with a Subset of the Ro Ribonucleoprotein-associated Y RNAs in Vitro and in Vivo. J. Biol. Chem.
276: 20711-20718
[Abstract]
[Full Text]