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

J Biol Chem. 1989 Dec 15;264(35):21183-9.

Abstract

Rearranged MOPC41 immunoglobulin kappa 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 kappa 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 kappa gene expression or only 6% with respect to the MOPC41 kappa 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Northern
  • Blotting, Southern
  • Cell Line
  • DNA, Neoplasm / genetics
  • Enhancer Elements, Genetic*
  • Gene Expression*
  • Genes, Immunoglobulin*
  • Immunoglobulin kappa-Chains / genetics*
  • Introns*
  • Mice
  • Plasmacytoma / genetics
  • Plasmacytoma / immunology
  • Plasmids
  • Restriction Mapping
  • Transfection*

Substances

  • DNA, Neoplasm
  • Immunoglobulin kappa-Chains