Transcription termination and chromatin structure of the active immunoglobulin κ gene locus

M. Xu, M. B. Barnard, S. M. Rose, P. N. Cockerill, S. Y. Huang, W. T. Garrard

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Abstract

To investigate the chromatin surrounding an active gene, we have determined the distribution of RNA polymerase molecules, the intactness of nucleosomal structure, and the subnuclear compartmentalization along 15 kilobase pairs (kb) of the mouse κ immunoglobulin locus of MPC-11 plasmacytoma cells. Hybridization of in vitro nuclear transcripts to probes specific for the template strand reveals that transcription terminates within the region between 1.1 and 2.3 kb downstream from the poly(A) addition site. Ten different short sequences (8-13 base pairs) reside within 460 base pairs of this termination region that exhibit homology with sequences found in the termination regions of mouse β-globin and chick ovalbumin genes. Transcription of the nontemplate strand occurs on either side of this termination region. We find that both within the transcription unit and 6.5 kb downstream of the termination region of the κ gene, the canonical nucleosomal structure is perturbed, the chromatin exhibits pronounced insolubility, and the nucleosomes liberated by micrococcal nuclease appear to lack histone H1. The insolubility is characterized by interactions that are disrupted by 0.3 to 0.6 M NaCl treatment. We conclude that the active chromatin phenotype spreads a considerable distance along the κ locus, well beyond the region of transcription termination.

Original languageEnglish (US)
Pages (from-to)3838-3845
Number of pages8
JournalJournal of Biological Chemistry
Volume261
Issue number8
Publication statusPublished - 1986

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ASJC Scopus subject areas

  • Biochemistry

Cite this

Xu, M., Barnard, M. B., Rose, S. M., Cockerill, P. N., Huang, S. Y., & Garrard, W. T. (1986). Transcription termination and chromatin structure of the active immunoglobulin κ gene locus. Journal of Biological Chemistry, 261(8), 3838-3845.