Immunoglobulin κ gene expression after stable integration. I. Role of the intronic MAR and enhancer in plasmacytoma cells

V. C. Blasquez, M. Xu, S. C. Moses, W. T. Garrard

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Abstract

Rearranged MOPC41 immunoglobulin κ gene constructs have been stably introduced into cultured S194 mouse plasmacytoma cells to investigate the effects of deleting the intronic enhancer and/or matrix association region (MAR) on gene expression. Intact single-copy κ genes containing 1.5 kilobase pairs of upstream and 8.5 kilobase pairs of downstream flanking sequences exhibited sensitivity to chromosome position effects and were expressed at a mean level of 27% relative to the endogenous κ gene expression or only 6% with respect to the MOPC41 κ mRNA levels in the tumor. Deletion of the intronic MAR led to a 4-fold decrease in expression, while deletion of both the MAR and enhancer led to an 11-fold decline. These effects were dampened by preselecting for integration into a transcriptionally poised chromatin location as demonstrated by linkage to a selectable marker which lacked both a MAR and an enhancer. Significantly, we found that sequences downstream of the poly(A) addition site compensated 150-fold for deletion of the intronic enhancer.

Original languageEnglish (US)
Pages (from-to)21183-21189
Number of pages7
JournalJournal of Biological Chemistry
Volume264
Issue number35
StatePublished - Dec 1 1989

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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