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Molecular and Cellular Biology, December 2003, p. 9338-9348, Vol. 23, No. 24
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.24.9338-9348.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The c-myc Insulator Element and Matrix Attachment Regions Define the c-myc Chromosomal Domain

Wendy M. Gombert,1 Stephen D. Farris,1 Eric D. Rubio,1 Kristin M. Morey-Rosler,1 William H. Schubach,2 and Anton Krumm1*

Department of Radiation Oncology, University of Washington School of Medicine, Seattle, Washington 98104,1 Division of Medical Oncology, Department of Medicine, Veterans Administration Puget Sound Health Care System, Seattle Division, Seattle, Washington 981082

Received 6 June 2003/ Returned for modification 6 August 2003/ Accepted 12 September 2003

Insulator elements and matrix attachment regions are essential for the organization of genetic information within the nucleus. By comparing the pattern of histone modifications at the mouse and human c-myc alleles, we identified an evolutionarily conserved boundary at which the c-myc transcription unit is separated from the flanking condensed chromatin enriched in lysine 9-methylated histone H3. This region harbors the c-myc insulator element (MINE), which contains at least two physically separable, functional activities: enhancer-blocking activity and barrier activity. The enhancer-blocking activity is mediated by CTCF. Chromatin immunoprecipitation assays demonstrate that CTCF is constitutively bound at the insulator and at the promoter region independent of the transcriptional status of c-myc. This result supports an architectural role of CTCF rather than a regulatory role in transcription. An additional higher-order nuclear organization of the c-myc locus is provided by matrix attachment regions (MARs) that define a domain larger than 160 kb. The MARs of the c-myc domain do not act to prevent the association of flanking regions with lysine 9-methylated histones, suggesting that they do not function as barrier elements.

* Corresponding author. Mailing address: VA Medical Center, 1660 S. Columbian Way R151, Seattle, WA 98108. Phone: (206) 764-2381. Fax: (206) 764-2827. E-mail: akrumm{at}u.washington.edu.

Molecular and Cellular Biology, December 2003, p. 9338-9348, Vol. 23, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.24.9338-9348.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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