Factors Binding a Non-classical Cis-element Prevent Heterochromatin Effects on Locus Control Region Activity

Faith Harrow, Jeanne U. Amuta, Shauna R. Hutchinson, Frank Akwaa, Benjamin D. Ortiz

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A locus control region (LCR) is a cis-acting gene-regulatory element capable of transferring the expression characteristics of its gene locus of origin to a linked transgene. Furthermore, it can do this independently of the site of integration in the genome of transgenic mice. Although most LCRs contain subelements with classical transcriptional enhancer function, key aspects of LCR activity are supported by cis-acting sequences devoid of the ability to act as direct transcriptional enhancers. Very few of these "non-enhancer" LCR components have been characterized. Consequently, the sequence requirements and molecular bases for their functions, as well as their roles in LCR activity, are poorly understood. We have investigated these questions using the LCR from the mouse T cell receptor (TCR) α/Dad1 gene locus. Here we focus on DNase hypersensitive site (HS) 6 of the TCRα LCR. HS6 does not support classical enhancer activity, yet has gene regulatory activity in an in vivo chromatin context. We have identified three in vivo occupied factor-binding sites within HS6, two of which interact with Runx1 and Elf-1 factors. Deletion of these sites from the LCR impairs its activity in vivo. This mutation renders the transgene locus abnormally inaccessible in chromatin, preventing the normal function of other LCR subelements and reducing transgene mRNA levels. These data show these factor-binding sites are required for preventing heterochromatin formation and indicate that they function to maintain an active TCRα LCR assembly in vivo.

Original languageEnglish (US)
Pages (from-to)17842-17849
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number17
DOIs
StatePublished - Apr 23 2004

Fingerprint

Locus Control Region
Heterochromatin
Genes
T-Cell Antigen Receptor
Transgenes
Regulator Genes
Chromatin
Binding Sites
T-Cell Receptor Genes
Deoxyribonucleases
Transgenic Mice
Genome

ASJC Scopus subject areas

  • Biochemistry

Cite this

Factors Binding a Non-classical Cis-element Prevent Heterochromatin Effects on Locus Control Region Activity. / Harrow, Faith; Amuta, Jeanne U.; Hutchinson, Shauna R.; Akwaa, Frank; Ortiz, Benjamin D.

In: Journal of Biological Chemistry, Vol. 279, No. 17, 23.04.2004, p. 17842-17849.

Research output: Contribution to journalArticle

Harrow, Faith ; Amuta, Jeanne U. ; Hutchinson, Shauna R. ; Akwaa, Frank ; Ortiz, Benjamin D. / Factors Binding a Non-classical Cis-element Prevent Heterochromatin Effects on Locus Control Region Activity. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 17. pp. 17842-17849.
@article{b9001f7443d54e36b8aed5fb9a9c404f,
title = "Factors Binding a Non-classical Cis-element Prevent Heterochromatin Effects on Locus Control Region Activity",
abstract = "A locus control region (LCR) is a cis-acting gene-regulatory element capable of transferring the expression characteristics of its gene locus of origin to a linked transgene. Furthermore, it can do this independently of the site of integration in the genome of transgenic mice. Although most LCRs contain subelements with classical transcriptional enhancer function, key aspects of LCR activity are supported by cis-acting sequences devoid of the ability to act as direct transcriptional enhancers. Very few of these {"}non-enhancer{"} LCR components have been characterized. Consequently, the sequence requirements and molecular bases for their functions, as well as their roles in LCR activity, are poorly understood. We have investigated these questions using the LCR from the mouse T cell receptor (TCR) α/Dad1 gene locus. Here we focus on DNase hypersensitive site (HS) 6 of the TCRα LCR. HS6 does not support classical enhancer activity, yet has gene regulatory activity in an in vivo chromatin context. We have identified three in vivo occupied factor-binding sites within HS6, two of which interact with Runx1 and Elf-1 factors. Deletion of these sites from the LCR impairs its activity in vivo. This mutation renders the transgene locus abnormally inaccessible in chromatin, preventing the normal function of other LCR subelements and reducing transgene mRNA levels. These data show these factor-binding sites are required for preventing heterochromatin formation and indicate that they function to maintain an active TCRα LCR assembly in vivo.",
author = "Faith Harrow and Amuta, {Jeanne U.} and Hutchinson, {Shauna R.} and Frank Akwaa and Ortiz, {Benjamin D.}",
year = "2004",
month = "4",
day = "23",
doi = "10.1074/jbc.M401258200",
language = "English (US)",
volume = "279",
pages = "17842--17849",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "17",

}

TY - JOUR

T1 - Factors Binding a Non-classical Cis-element Prevent Heterochromatin Effects on Locus Control Region Activity

AU - Harrow, Faith

AU - Amuta, Jeanne U.

AU - Hutchinson, Shauna R.

AU - Akwaa, Frank

AU - Ortiz, Benjamin D.

PY - 2004/4/23

Y1 - 2004/4/23

N2 - A locus control region (LCR) is a cis-acting gene-regulatory element capable of transferring the expression characteristics of its gene locus of origin to a linked transgene. Furthermore, it can do this independently of the site of integration in the genome of transgenic mice. Although most LCRs contain subelements with classical transcriptional enhancer function, key aspects of LCR activity are supported by cis-acting sequences devoid of the ability to act as direct transcriptional enhancers. Very few of these "non-enhancer" LCR components have been characterized. Consequently, the sequence requirements and molecular bases for their functions, as well as their roles in LCR activity, are poorly understood. We have investigated these questions using the LCR from the mouse T cell receptor (TCR) α/Dad1 gene locus. Here we focus on DNase hypersensitive site (HS) 6 of the TCRα LCR. HS6 does not support classical enhancer activity, yet has gene regulatory activity in an in vivo chromatin context. We have identified three in vivo occupied factor-binding sites within HS6, two of which interact with Runx1 and Elf-1 factors. Deletion of these sites from the LCR impairs its activity in vivo. This mutation renders the transgene locus abnormally inaccessible in chromatin, preventing the normal function of other LCR subelements and reducing transgene mRNA levels. These data show these factor-binding sites are required for preventing heterochromatin formation and indicate that they function to maintain an active TCRα LCR assembly in vivo.

AB - A locus control region (LCR) is a cis-acting gene-regulatory element capable of transferring the expression characteristics of its gene locus of origin to a linked transgene. Furthermore, it can do this independently of the site of integration in the genome of transgenic mice. Although most LCRs contain subelements with classical transcriptional enhancer function, key aspects of LCR activity are supported by cis-acting sequences devoid of the ability to act as direct transcriptional enhancers. Very few of these "non-enhancer" LCR components have been characterized. Consequently, the sequence requirements and molecular bases for their functions, as well as their roles in LCR activity, are poorly understood. We have investigated these questions using the LCR from the mouse T cell receptor (TCR) α/Dad1 gene locus. Here we focus on DNase hypersensitive site (HS) 6 of the TCRα LCR. HS6 does not support classical enhancer activity, yet has gene regulatory activity in an in vivo chromatin context. We have identified three in vivo occupied factor-binding sites within HS6, two of which interact with Runx1 and Elf-1 factors. Deletion of these sites from the LCR impairs its activity in vivo. This mutation renders the transgene locus abnormally inaccessible in chromatin, preventing the normal function of other LCR subelements and reducing transgene mRNA levels. These data show these factor-binding sites are required for preventing heterochromatin formation and indicate that they function to maintain an active TCRα LCR assembly in vivo.

UR - http://www.scopus.com/inward/record.url?scp=2342441326&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2342441326&partnerID=8YFLogxK

U2 - 10.1074/jbc.M401258200

DO - 10.1074/jbc.M401258200

M3 - Article

C2 - 14966120

AN - SCOPUS:2342441326

VL - 279

SP - 17842

EP - 17849

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 17

ER -