Nucleic Acids Research, 1994, Vol. 22, No. 3 370-375
© 1994
MOLECULAR BIOLOGY |
Micrococcal nuclease digestion of nuclei reveals extended nucleosome ladders having anomalous DNA lengths for chromatin assembled on non-replicating plasmids in transfected cells
Department of Biological Sciences Purdue University, West Lafayette, IN 47907, USA
*To whom correspondence should be addressed
Received October 5, 1993. Accepted December 22, 1993.
The chromatin structures of a variety of plasmids and plasmid constructions, transiently transfected into mouse Ltk– cells using the DEAE-dextran procedure, were studied by micrococcal nuclease digestion of nuclei and Southern hybridization. Although regularly arranged nucleosome-like particles dearly were formed on the transfected DNA, the nucleosome ladders, in some cases with 13–14 bands, were anomalous. Most often, a ladder of DNA fragments with lengths of approximately 300, 500, 700, 900 bp, etc. was generated. In contrast, typical 180–190 bp multiples were generated from bulk cellular or endogenous ß-actin gene chromatin. Very similar results were obtained with all DNA's transfected, and in a variety of cell lines, provided that plasmid replication did not occur. Additionally, after digestion of nuclei, about 90% of the chromatin fragments that contained transfected DNA sequences could not be solubilized at low ionic strength, in contrast with bulk cellular chromatin, suggesting association with nuclear structures or nuclear matrix. The remaining 10% of transfected DNA sequences, arising from soluble chromatin fragments, generated a typical nucleosome ladder. These results are consistent with the idea that assembly of atypical chromatin structures might be induced by proximity to elements of the nuclear pore complex or by nuclear compartmentallzation.
+Presertt address: Fred Hutchinson Cancer Research Center, Basic Sciences Division, Seattle, WA 98104, USA
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  P. B. Hebbar and T. K. Archer Altered Histone H1 Stoichiometry and an Absence of Nucleosome Positioning on Transfected DNA J. Biol. Chem., February 22, 2008; 283(8): 4595 - 4601. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. L. Bishop, M. Ramalho, N. Nadkarni, W. May Kong, C. F. Higgins, and N. Krauzewicz Role for Centromeric Heterochromatin and PML Nuclear Bodies in the Cellular Response to Foreign DNA. Mol. Cell. Biol., April 1, 2006; 26(7): 2583 - 2594. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D. Stewart, J. Li, and J. Wong Relationship between Histone H3 Lysine 9 Methylation, Transcription Repression, and Heterochromatin Protein 1 Recruitment Mol. Cell. Biol., April 1, 2005; 25(7): 2525 - 2538. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Wang and J. Zhu The hTERT Gene Is Embedded in a Nuclease-resistant Chromatin Domain J. Biol. Chem., December 31, 2004; 279(53): 55401 - 55410. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Ferrari, K. C. Simmen, Y. Dusserre, K. Muller, G. Fourel, E. Gilson, and N. Mermod Chromatin Domain Boundaries Delimited by a Histone-binding Protein in Yeast J. Biol. Chem., December 31, 2004; 279(53): 55520 - 55530. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Krieg, S. A. Krieg, B. S. Ahn, and D. J. Shapiro Interplay between Estrogen Response Element Sequence and Ligands Controls in Vivo Binding of Estrogen Receptor to Regulated Genes J. Biol. Chem., February 6, 2004; 279(6): 5025 - 5034. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Wang and J. Zhu Evidence for a Relief of Repression Mechanism for Activation of the Human Telomerase Reverse Transcriptase Promoter J. Biol. Chem., May 23, 2003; 278(21): 18842 - 18850. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Matsui, H. A. Chansky, F. Barahmand-Pour, A. Zielinska-Kwiatkowska, N. Tsumaki, A. Myoui, H. Yoshikawa, L. Yang, and D. R. Eyre COL11A2 Collagen Gene Transcription Is Differentially Regulated by EWS/ERG Sarcoma Fusion Protein and Wild-type ERG J. Biol. Chem., March 21, 2003; 278(13): 11369 - 11375. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Marin-Husstege, M. Muggironi, A. Liu, and P. Casaccia-Bonnefil Histone Deacetylase Activity Is Necessary for Oligodendrocyte Lineage Progression J. Neurosci., December 1, 2002; 22(23): 10333 - 10345. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-Y. Xie, V. P. Bermudez, and W. R. Folk Stimulation of DNA Replication from the Polyomavirus Origin by PCAF and GCN5 Acetyltransferases: Acetylation of Large T Antigen Mol. Cell. Biol., November 15, 2002; 22(22): 7907 - 7918. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Quivy, E. Adam, Y. Collette, D. Demonte, A. Chariot, C. Vanhulle, B. Berkhout, R. Castellano, Y. de Launoit, A. Burny, et al. Synergistic Activation of Human Immunodeficiency Virus Type 1 Promoter Activity by NF-{kappa}B and Inhibitors of Deacetylases: Potential Perspectives for the Development of Therapeutic Strategies J. Virol., October 2, 2002; 76(21): 11091 - 11103. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. P. Ashburner, S. D. Westerheide, and A. S. Baldwin Jr. The p65 (RelA) Subunit of NF-{kappa}B Interacts with the Histone Deacetylase (HDAC) Corepressors HDAC1 and HDAC2 To Negatively Regulate Gene Expression Mol. Cell. Biol., October 15, 2001; 21(20): 7065 - 7077. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Cakouros, P. N. Cockerill, A. G. Bert, R. Mital, D. C. Roberts, and M. F. Shannon A NF-{{kappa}}B/Sp1 Region Is Essential for Chromatin Remodeling and Correct Transcription of a Human Granulocyte- Macrophage Colony-Stimulating Factor Transgene J. Immunol., July 1, 2001; 167(1): 302 - 310. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. C. Harnish, M. S. Scicchitano, S. J. Adelman, C. R. Lyttle, and S. K. Karathanasis The Role of CBP in Estrogen Receptor Cross-Talk with Nuclear Factor-{kappa}B in HepG2 Cells Endocrinology, September 1, 2000; 141(9): 3403 - 3411. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. JAFFUEL, P. DEMOLY, C. GOUGAT, P. BALAGUER, G. MAUTINO, P. GODARD, J. BOUSQUET, and M. MATHIEU Transcriptional Potencies of Inhaled Glucocorticoids Am. J. Respir. Crit. Care Med., July 1, 2000; 162(1): 57 - 63. [Abstract] [Full Text]
|
|
|
|
|
|
C. Mao and D. J. Shapiro A Histone Deacetylase Inhibitor Potentiates Estrogen Receptor Activation of a Stably Integrated Vitellogenin Promoter in HepG2 Cells Endocrinology, July 1, 2000; 141(7): 2361 - 2369. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Illi, P. Puri, L. Morgante, M. C. Capogrossi, and C. Gaetano Nuclear Factor-{kappa}B and cAMP Response Element Binding Protein Mediate Opposite Transcriptional Effects on the Flk-1/KDR Gene Promoter Circ. Res., June 23, 2000; 86 (12): e110 - e117. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. R. Dorris and K. Struhl Artificial Recruitment of TFIID, but Not RNA Polymerase II Holoenzyme, Activates Transcription in Mammalian Cells Mol. Cell. Biol., June 15, 2000; 20(12): 4350 - 4358. [Abstract] [Full Text]
|
|
|
|
|
|
L. F. Michael, H. Asahara, A. I. Shulman, W. L. Kraus, and M. Montminy The Phosphorylation Status of a Cyclic AMP-Responsive Activator Is Modulated via a Chromatin-Dependent Mechanism Mol. Cell. Biol., March 1, 2000; 20(5): 1596 - 1603. [Abstract] [Full Text]
|
|
|
|
 |
|
 N. E. Wittekindt, K. Hortnagel, C. Geltinger, and A. Polack Activation of c-myc promoter P1 by immunoglobulin {kappa} gene enhancers in Burkitt lymphoma: functional characterization of the intron enhancer motifs {kappa}B, E box 1 and E box 2, and of the 3' enhancer motif PU Nucleic Acids Res., February 1, 2000; 28(3): 800 - 808. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Ito, Y. Park, J. Weck, K. E. Mayo, and J. L. Jameson Synergistic Activation of the Inhibin {alpha}-Promoter by Steroidogenic Factor-1 and Cyclic Adenosine 3',5'-Monophosphate Mol. Endocrinol., January 1, 2000; 14(1): 66 - 81. [Abstract] [Full Text]
|
|
|
|
 |
|
 F. Recillas-Targa, A. C. Bell, and G. Felsenfeld Positional enhancer-blocking activity of the chicken beta -globin insulator in transiently transfected cells PNAS, December 7, 1999; 96(25): 14354 - 14359. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. J. Fry, A. Pearson, E. Malinowski, S. M. Bartley, J. Greenblatt, and P. J. Farnham Activation of the Murine Dihydrofolate Reductase Promoter by E2F1. A REQUIREMENT FOR CBP RECRUITMENT J. Biol. Chem., May 28, 1999; 274(22): 15883 - 15891. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Espinos, A. L. V. Thai, C. Pomies, and M. J. Weber Cooperation between Phosphorylation and Acetylation Processes in Transcriptional Control Mol. Cell. Biol., May 1, 1999; 19(5): 3474 - 3484. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Zanta, P. Belguise-Valladier, and J.-P. Behr Gene delivery: A single nuclear localization signal peptide is sufficient to carry DNA to the cell nucleus PNAS, January 5, 1999; 96(1): 91 - 96. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. D. Bailey, T. Pavelitz, and A. M. Weiner The Microsatellite Sequence (CT)n · (GA)n Promotes Stable Chromosomal Integration of Large Tandem Arrays of Functional Human U2 Small Nuclear RNA Genes Mol. Cell. Biol., April 1, 1998; 18(4): 2262 - 2271. [Abstract] [Full Text]
|
|
|
|
|
|
G. Fourel, F. Ringeisen, M. Flajolet, P. Tiollais, and M. A. Buendia Functional Analysis of Ground Squirrel Hepatitis Virus Enhancer II J. Virol., February 1, 1998; 72(2): 1616 - 1622. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. L. Smith and G. L. Hager Transcriptional Regulation of Mammalian Genes in Vivo. A TALE OF TWO TEMPLATES J. Biol. Chem., October 31, 1997; 272(44): 27493 - 27496. [Full Text] [PDF]
|
|
|
|
|
|
S. Boronat, H. Richard-Foy, and B. Pina Specific Deactivation of the Mouse Mammary Tumor Virus Long Terminal Repeat Promoter upon Continuous Hormone Treatment J. Biol. Chem., August 29, 1997; 272(35): 21803 - 21810. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Jenster, T. E. Spencer, M. M. Burcin, S. Y. Tsai, M.-J. Tsai, and B. W. O'Malley Steroid receptor induction of gene transcription: A two-step model PNAS, July 22, 1997; 94(15): 7879 - 7884. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A N Gerber, T R Klesert, D A Bergstrom, and S J Tapscott Two domains of MyoD mediate transcriptional activation of genes in repressive chromatin: a mechanism for lineage determination in myogenesis. Genes & Dev., February 15, 1997; 11(4): 436 - 450. [Abstract] [PDF]
|
|
|
|
|
|
K. K. Bhakat and S. Mitra Regulation of the Human O6-Methylguanine-DNA Methyltransferase Gene by Transcriptional Coactivators cAMP Response Element-binding Protein-binding Protein and p300 J. Biol. Chem., October 27, 2000; 275(44): 34197 - 34204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. D. Urnov and A. P. Wolffe An Array of Positioned Nucleosomes Potentiates Thyroid Hormone Receptor Action in Vivo J. Biol. Chem., June 1, 2001; 276(23): 19753 - 19761. [Abstract] [Full Text] [PDF]
|
|