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

Research output: Contribution to journalArticle

96 Citations (Scopus)

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 - 1989

Fingerprint

Immunoglobulin Genes
Plasmacytoma
Gene expression
Immunoglobulins
Gene Expression
Poly A
Chromatin
Genes
Chromosomes
Messenger RNA
Tumors
Neoplasms

ASJC Scopus subject areas

  • Biochemistry

Cite this

Immunoglobulin κ gene expression after stable integration. I. Role of the intronic MAR and enhancer in plasmacytoma cells. / Blasquez, V. C.; Xu, M.; Moses, S. C.; Garrard, W. T.

In: Journal of Biological Chemistry, Vol. 264, No. 35, 1989, p. 21183-21189.

Research output: Contribution to journalArticle

Blasquez, V. C. ; Xu, M. ; Moses, S. C. ; Garrard, W. T. / Immunoglobulin κ gene expression after stable integration. I. Role of the intronic MAR and enhancer in plasmacytoma cells. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 35. pp. 21183-21189.
@article{7fe27eb0f1434917b589f08e47c86d43,
title = "Immunoglobulin κ gene expression after stable integration. I. Role of the intronic MAR and enhancer in plasmacytoma cells",
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.",
author = "Blasquez, {V. C.} and M. Xu and Moses, {S. C.} and Garrard, {W. T.}",
year = "1989",
language = "English (US)",
volume = "264",
pages = "21183--21189",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "35",

}

TY - JOUR

T1 - Immunoglobulin κ gene expression after stable integration. I. Role of the intronic MAR and enhancer in plasmacytoma cells

AU - Blasquez, V. C.

AU - Xu, M.

AU - Moses, S. C.

AU - Garrard, W. T.

PY - 1989

Y1 - 1989

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0024846566&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024846566&partnerID=8YFLogxK

M3 - Article

VL - 264

SP - 21183

EP - 21189

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 35

ER -