A targeted κ immunoglobulin gene containing a deletion of the nuclear matrix association region exhibits spontaneous hyper-recombination in pre-B cells

Michael A. Hale, William T. Garrard

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Previous studies employing ectopic integration of reporter genes have shown that the nuclear matrix association region (MAR) adjacent to the intronic enhancer of the mouse κ immunoglobulin (Ig) gene is required for high level transcription of rearranged genes, demethylation, reduction of position effects and maximal somatic hypermutation in B cells. To test for the function of this MAR in its natural chromosomal environment, we pursued the 'HIT-and-RUN' procedure with the mouse pre-B cell line 103 to create a targeted MAR deletion. We observed a 'HIT' targeting frequency of 1/684 but 0/2100 'RUN' clones maintained the MAR-deleted germline locus because of an unexpected hyper-recombination for Vκ-Jκ joining, specifically to the MAR- deleted allele, and primarily at Jκ4 and Jκ5. This hyper-recombination was correlated with undermethylation of the Jκ-Cκ region but not with the level of local transcription. These results are consistent with the possibility that the MAR and/or DNA methylation negatively regulate(s) Vκ-Jκ joining during the pre-B cell stage of development.

Original languageEnglish (US)
Pages (from-to)609-620
Number of pages12
JournalMolecular Immunology
Volume35
Issue number10
DOIs
StatePublished - Jul 15 1998

Fingerprint

Nuclear Matrix
Immunoglobulin Genes
B-Lymphoid Precursor Cells
Genetic Recombination
DNA Methylation
Reporter Genes
B-Lymphocytes
Clone Cells
Alleles
Cell Line
Genes

Keywords

  • κ immunoglobulin gene
  • Gene rearrangement
  • MAR
  • V- and J-region usage

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

A targeted κ immunoglobulin gene containing a deletion of the nuclear matrix association region exhibits spontaneous hyper-recombination in pre-B cells. / Hale, Michael A.; Garrard, William T.

In: Molecular Immunology, Vol. 35, No. 10, 15.07.1998, p. 609-620.

Research output: Contribution to journalArticle

@article{f5e91d93de84419e8fea51f81a5ca6c6,
title = "A targeted κ immunoglobulin gene containing a deletion of the nuclear matrix association region exhibits spontaneous hyper-recombination in pre-B cells",
abstract = "Previous studies employing ectopic integration of reporter genes have shown that the nuclear matrix association region (MAR) adjacent to the intronic enhancer of the mouse κ immunoglobulin (Ig) gene is required for high level transcription of rearranged genes, demethylation, reduction of position effects and maximal somatic hypermutation in B cells. To test for the function of this MAR in its natural chromosomal environment, we pursued the 'HIT-and-RUN' procedure with the mouse pre-B cell line 103 to create a targeted MAR deletion. We observed a 'HIT' targeting frequency of 1/684 but 0/2100 'RUN' clones maintained the MAR-deleted germline locus because of an unexpected hyper-recombination for Vκ-Jκ joining, specifically to the MAR- deleted allele, and primarily at Jκ4 and Jκ5. This hyper-recombination was correlated with undermethylation of the Jκ-Cκ region but not with the level of local transcription. These results are consistent with the possibility that the MAR and/or DNA methylation negatively regulate(s) Vκ-Jκ joining during the pre-B cell stage of development.",
keywords = "κ immunoglobulin gene, Gene rearrangement, MAR, V- and J-region usage",
author = "Hale, {Michael A.} and Garrard, {William T.}",
year = "1998",
month = "7",
day = "15",
doi = "10.1016/S0161-5890(98)00060-1",
language = "English (US)",
volume = "35",
pages = "609--620",
journal = "Molecular Immunology",
issn = "0161-5890",
publisher = "Elsevier Limited",
number = "10",

}

TY - JOUR

T1 - A targeted κ immunoglobulin gene containing a deletion of the nuclear matrix association region exhibits spontaneous hyper-recombination in pre-B cells

AU - Hale, Michael A.

AU - Garrard, William T.

PY - 1998/7/15

Y1 - 1998/7/15

N2 - Previous studies employing ectopic integration of reporter genes have shown that the nuclear matrix association region (MAR) adjacent to the intronic enhancer of the mouse κ immunoglobulin (Ig) gene is required for high level transcription of rearranged genes, demethylation, reduction of position effects and maximal somatic hypermutation in B cells. To test for the function of this MAR in its natural chromosomal environment, we pursued the 'HIT-and-RUN' procedure with the mouse pre-B cell line 103 to create a targeted MAR deletion. We observed a 'HIT' targeting frequency of 1/684 but 0/2100 'RUN' clones maintained the MAR-deleted germline locus because of an unexpected hyper-recombination for Vκ-Jκ joining, specifically to the MAR- deleted allele, and primarily at Jκ4 and Jκ5. This hyper-recombination was correlated with undermethylation of the Jκ-Cκ region but not with the level of local transcription. These results are consistent with the possibility that the MAR and/or DNA methylation negatively regulate(s) Vκ-Jκ joining during the pre-B cell stage of development.

AB - Previous studies employing ectopic integration of reporter genes have shown that the nuclear matrix association region (MAR) adjacent to the intronic enhancer of the mouse κ immunoglobulin (Ig) gene is required for high level transcription of rearranged genes, demethylation, reduction of position effects and maximal somatic hypermutation in B cells. To test for the function of this MAR in its natural chromosomal environment, we pursued the 'HIT-and-RUN' procedure with the mouse pre-B cell line 103 to create a targeted MAR deletion. We observed a 'HIT' targeting frequency of 1/684 but 0/2100 'RUN' clones maintained the MAR-deleted germline locus because of an unexpected hyper-recombination for Vκ-Jκ joining, specifically to the MAR- deleted allele, and primarily at Jκ4 and Jκ5. This hyper-recombination was correlated with undermethylation of the Jκ-Cκ region but not with the level of local transcription. These results are consistent with the possibility that the MAR and/or DNA methylation negatively regulate(s) Vκ-Jκ joining during the pre-B cell stage of development.

KW - κ immunoglobulin gene

KW - Gene rearrangement

KW - MAR

KW - V- and J-region usage

UR - http://www.scopus.com/inward/record.url?scp=0031790033&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031790033&partnerID=8YFLogxK

U2 - 10.1016/S0161-5890(98)00060-1

DO - 10.1016/S0161-5890(98)00060-1

M3 - Article

C2 - 9823759

AN - SCOPUS:0031790033

VL - 35

SP - 609

EP - 620

JO - Molecular Immunology

JF - Molecular Immunology

SN - 0161-5890

IS - 10

ER -