![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 9, Issue 6, 1523-1536, June 1998
Sir William Dunn School of Pathology, University of Oxford, Oxford,
OX1 3RE, United Kingdom
Using HeLa cells, we have developed methods to determine 1) the
number of RNA polymerases that are active at any moment, 2) the number
of transcription sites, and 3) the number of polymerases associated
with one transcription unit. To count engaged polymerases, cells were
encapsulated in agarose, permeabilized, treated with ribonuclease, and
the now-truncated transcripts extended in [32P]uridine
triphosphate; then, the number of growing transcripts was calculated
from the total number of nucleotides incorporated and the average
increment in length of the transcripts. Approximately 15,000 transcripts were elongated by polymerase I, and ~75,000 were
elongated by polymerases II and III. Transcription sites were detected
after the cells were grown in bromouridine for <2.5 min, after which
the resulting bromo-RNA was labeled with gold particles; electron
microscopy showed that most extranucleolar transcripts were
concentrated in ~2400 sites with diameters of ~80 nm. The number of
polymerases associated with a transcription unit was counted after
templates were spread over a large area; most extranucleolar units were
associated with one elongating complex. These results suggest that many
templates are attached in a "cloud" of loops around a site; each
site, or transcription "factory," would contain ~30 active
polymerases and associated transcripts.
This article has been cited by other articles:
|
|
 |
|
  C. H. Eskiw, A. Rapp, D. R. F. Carter, and P. R. Cook RNA polymerase II activity is located on the surface of protein-rich transcription factories J. Cell Sci., June 15, 2008; 121(12): 1999 - 2007. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Vecchio, L. Solimando, M. Biggiogera, and S. Fakan Use of Halogenated Precursors for Simultaneous DNA and RNA Detection by Means of Immunoelectron and Immunofluorescence Microscopy J. Histochem. Cytochem., January 1, 2008; 56(1): 45 - 55. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Schneider and R. Grosschedl Dynamics and interplay of nuclear architecture, genome organization, and gene expression Genes & Dev., December 1, 2007; 21(23): 3027 - 3043. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Binnie, P. Castelo-Branco, J. Monks, and N. J. Proudfoot Homologous gene sequences mediate transcription-domain formation J. Cell Sci., September 15, 2006; 119(18): 3876 - 3887. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Bon, S. J. McGowan, and P. R. Cook Many expressed genes in bacteria and yeast are transcribed only once per cell cycle FASEB J, August 1, 2006; 20(10): 1721 - 1723. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G.-L. Zhou, L. Xin, W. Song, L.-J. Di, G. Liu, X.-S. Wu, D.-P. Liu, and C.-C. Liang Active Chromatin Hub of the Mouse {alpha}-Globin Locus Forms in a Transcription Factory of Clustered Housekeeping Genes. Mol. Cell. Biol., July 1, 2006; 26(13): 5096 - 5105. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. S. Dimitrova Nuclear transcription is essential for specification of mammalian replication origins. Genes Cells, July 1, 2006; 11(7): 829 - 844. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Faro-Trindade and P. R. Cook A Conserved Organization of Transcription during Embryonic Stem Cell Differentiation and in Cells with High C Value Mol. Biol. Cell, July 1, 2006; 17(7): 2910 - 2920. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Fraser and J. D. Engel Constricting restricted transcription: the (actively?) shrinking web Genes & Dev., June 1, 2006; 20(11): 1379 - 1383. [Full Text] [PDF]
|
|
|
|
 |
|
 D. Marenduzzo, C. Micheletti, and P. R. Cook Entropy-Driven Genome Organization Biophys. J., May 15, 2006; 90(10): 3712 - 3721. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Q. Xie, S. Martin, P. V. Guillot, D. L. Bentley, and A. Pombo Splicing Speckles Are Not Reservoirs of RNA Polymerase II, but Contain an Inactive Form, Phosphorylated on Serine2 Residues of the C-Terminal Domain Mol. Biol. Cell, April 1, 2006; 17(4): 1723 - 1733. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. G. Thomas, L. J. M. Tosar, M. Loschi, J. M. Pasquini, J. Correale, S. Kindler, and G. L. Boccaccio Staufen Recruitment into Stress Granules Does Not Affect Early mRNA Transport in Oligodendrocytes Mol. Biol. Cell, January 1, 2005; 16(1): 405 - 420. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Martin, A. V. Failla, U. Spori, C. Cremer, and A. Pombo Measuring the Size of Biological Nanostructures with Spatially Modulated Illumination Microscopy Mol. Biol. Cell, May 1, 2004; 15(5): 2449 - 2455. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. French, Y. N. Osheim, F. Cioci, M. Nomura, and A. L. Beyer In Exponentially Growing Saccharomyces cerevisiae Cells, rRNA Synthesis Is Determined by the Summed RNA Polymerase I Loading Rate Rather than by the Number of Active Genes Mol. Cell. Biol., March 1, 2003; 23(5): 1558 - 1568. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Kimura, K. Sugaya, and P. R. Cook The transcription cycle of RNA polymerase II in living cells J. Cell Biol., December 9, 2002; 159(5): 777 - 782. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Koberna, J. Malinsky, A. Pliss, M. Masata, J. Vecerova, M. Fialova, J. Bednar, and I. Raska Ribosomal genes in focus: new transcripts label the dense fibrillar components and form clusters indicative of "Christmas trees" in situ J. Cell Biol., May 28, 2002; 157(5): 743 - 748. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. M. Gasser Visualizing Chromatin Dynamics in Interphase Nuclei Science, May 24, 2002; 296(5572): 1412 - 1416. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Snaar, P. Verdijk, H. J. Tanke, and R. W. Dirks Kinetics of HCMV immediate early mRNA expression in stably transfected fibroblasts J. Cell Sci., January 15, 2002; 115(2): 321 - 328. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. BURNS, D. A. JACKSON, and A. B. HASSAN A view through the clouds of imprinting FASEB J, August 1, 2001; 15(10): 1694 - 1703. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Caron, B. v. Schaik, M. v. d. Mee, F. Baas, G. Riggins, P. v. Sluis, M.-C. Hermus, R. v. Asperen, K. Boon, P. A. Voûte, et al. The Human Transcriptome Map: Clustering of Highly Expressed Genes in Chromosomal Domains Science, February 16, 2001; 291(5507): 1289 - 1292. [Abstract] [Full Text]
|
|
|
|
|
|
J Nickerson Experimental observations of a nuclear matrix J. Cell Sci., January 2, 2001; 114(3): 463 - 474. [Abstract] [PDF]
|
|
|
|
|
|
S. A. Enkemann, R. D. Ward, and S. L. Berger Mobility Within the Nucleus and Neighboring Cytosol Is a Key Feature of Prothymosin-{alpha} J. Histochem. Cytochem., October 1, 2000; 48(10): 1341 - 1356. [Abstract] [Full Text]
|
|
|
|
|
|
J. D. Lewis and D. Tollervey Like Attracts Like: Getting RNA Processing Together in the Nucleus Science, May 26, 2000; 288(5470): 1385 - 1389. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. N. Szentirmay and M. Sawadogo SURVEY AND SUMMARY: Spatial organization of RNA polymerase II transcription in the nucleus Nucleic Acids Res., May 15, 2000; 28(10): 2019 - 2025. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. L. Stenoien, M. G. Mancini, K. Patel, E. A. Allegretto*, C. L. Smith, and M. A. Mancini Subnuclear Trafficking of Estrogen Receptor-{alpha} and Steroid Receptor Coactivator-1 Mol. Endocrinol., April 1, 2000; 14(4): 518 - 534. [Abstract] [Full Text]
|
|
|
|
|
|
D. A. JACKSON, A. POMBO, and F. IBORRA The balance sheet for transcription: an analysis of nuclear RNA metabolism in mammalian cells FASEB J, February 1, 2000; 14(2): 242 - 254. [Abstract] [Full Text]
|
|
|
|
|
|
K Sugaya, M Vigneron, and P. Cook Mammalian cell lines expressing functional RNA polymerase II tagged with the green fluorescent protein J. Cell Sci., January 8, 2000; 113(15): 2679 - 2683. [Abstract] [PDF]
|
|
|
|
|
|
X. Wei, S. Somanathan, J. Samarabandu, and R. Berezney Three-dimensional Visualization of Transcription Sites and Their Association with Splicing Factor–rich Nuclear Speckles J. Cell Biol., August 9, 1999; 146(3): 543 - 558. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kimura, Y. Tao, R. G. Roeder, and P. R. Cook Quantitation of RNA Polymerase II and Its Transcription Factors in an HeLa Cell: Little Soluble Holoenzyme but Significant Amounts of Polymerases Attached to the Nuclear Substructure Mol. Cell. Biol., August 1, 1999; 19(8): 5383 - 5392. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. R. Cook The Organization of Replication and Transcription Science, June 11, 1999; 284(5421): 1790 - 1795. [Abstract] [Full Text]
|
|
|
|
|
|
A. Pombo, M. Hollinshead, and P. R. Cook Bridging the Resolution Gap: Imaging the Same Transcription Factories in Cryosections by Light and Electron Microscopy J. Histochem. Cytochem., April 1, 1999; 47(4): 471 - 480. [Abstract] [Full Text]
|
|
|
|
|
|
S. Panse, C Masson, L Heliot, J. Chassery, H. Junera, and D Hernandez-Verdun 3-D organization of ribosomal transcription units after DRB inhibition of RNA polymerase II transcription J. Cell Sci., January 7, 1999; 112(13): 2145 - 2154. [Abstract] [PDF]
|
|
|
|
|
|
D. Cmarko, P. J. Verschure, T. E. Martin, M. E. Dahmus, S. Krause, X.-D. Fu, R. van Driel, and S. Fakan Ultrastructural Analysis of Transcription and Splicing in the Cell Nucleus after Bromo-UTP Microinjection Mol. Biol. Cell, January 1, 1999; 10(1): 211 - 223. [Abstract] [Full Text]
|
|
|
|
|
|
T. Misteli, J. F. Caceres, J. Q. Clement, A. R. Krainer, M. F. Wilkinson, and D. L. Spector Serine Phosphorylation of SR Proteins Is Required for Their Recruitment to Sites of Transcription In Vivo J. Cell Biol., October 19, 1998; 143(2): 297 - 307. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Hendzel, M. J. Kruhlak, and D. P. Bazett-Jones Organization of Highly Acetylated Chromatin around Sites of Heterogeneous Nuclear RNA Accumulation Mol. Biol. Cell, September 1, 1998; 9(9): 2491 - 2507. [Abstract] [Full Text]
|
|
|
|
|
|
F. Iborra, D. Jackson, and P. Cook The path of transcripts from extra-nucleolar synthetic sites to nuclear pores: transcripts in transit are concentrated in discrete structures containing SR proteins J. Cell Sci., January 8, 1998; 111(15): 2269 - 2282. [Abstract] [PDF]
|
|
