Combinatorial regulation of the murine RAG-2 promoter by Sp1 and distinct lymphocyte-specific transcription factors

Gustavo A. Miranda, Maria Villalvazo, Zoran Galic, Jackelyn Alva, Roxanna Abrines, Yvette Yates, Cory J. Evans, Renato J. Aguilera

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The recombination activation genes, RAG-1 and RAG-2, encode the critical components of the recombinase complex responsible for the generation of functional antigen receptor genes. In order to gain an insight into the transcription factors and cis-acting elements that regulate the lymphocyte-specific expression of RAG-2, the promoter-region of this gene was isolated and characterized. This analysis demonstrated that a relatively small promoter fragment could confer lymphocyte-restricted expression to a reporter construct. Our work and that of others subsequently revealed that RAG-2 promoter expression is positively regulated by BSAP (PAX-5) and c-Myb transcription factors in B- and T-lineage cells, respectively. Although BSAP and c-Myb were deemed necessary for lymphocyte-specific expression, our analysis also uncovered a G-rich region at the 5′-end of the core promoter that was essential for full activity in lymphocyte cell lines. Site-directed mutagenesis revealed that a GA-box within the G-rich region was required for full promoter activity and subsequent DNA binding assays demonstrated that this element was specifically recognized by Sp1. Apart from showing that Sp1 interacts within the RAG-2 promoter, we also demonstrate that the Sp1-binding site is necessary for the high-level activation of this promoter.

Original languageEnglish (US)
Pages (from-to)1151-1159
Number of pages9
JournalMolecular Immunology
Volume38
Issue number15
DOIs
StatePublished - Jun 19 2002

Fingerprint

Transcription Factors
Lymphocytes
RAG-1 Genes
Recombinases
Antigen Receptors
Site-Directed Mutagenesis
Genetic Promoter Regions
Genetic Recombination
Genes
Binding Sites
Cell Line
DNA

All Science Journal Classification (ASJC) codes

  • Immunology
  • Molecular Biology

Cite this

Combinatorial regulation of the murine RAG-2 promoter by Sp1 and distinct lymphocyte-specific transcription factors. / Miranda, Gustavo A.; Villalvazo, Maria; Galic, Zoran; Alva, Jackelyn; Abrines, Roxanna; Yates, Yvette; Evans, Cory J.; Aguilera, Renato J.

In: Molecular Immunology, Vol. 38, No. 15, 19.06.2002, p. 1151-1159.

Research output: Contribution to journalArticle

Miranda, Gustavo A. ; Villalvazo, Maria ; Galic, Zoran ; Alva, Jackelyn ; Abrines, Roxanna ; Yates, Yvette ; Evans, Cory J. ; Aguilera, Renato J. / Combinatorial regulation of the murine RAG-2 promoter by Sp1 and distinct lymphocyte-specific transcription factors. In: Molecular Immunology. 2002 ; Vol. 38, No. 15. pp. 1151-1159.
@article{51c715f6df754c54a66254acfe073a6b,
title = "Combinatorial regulation of the murine RAG-2 promoter by Sp1 and distinct lymphocyte-specific transcription factors",
abstract = "The recombination activation genes, RAG-1 and RAG-2, encode the critical components of the recombinase complex responsible for the generation of functional antigen receptor genes. In order to gain an insight into the transcription factors and cis-acting elements that regulate the lymphocyte-specific expression of RAG-2, the promoter-region of this gene was isolated and characterized. This analysis demonstrated that a relatively small promoter fragment could confer lymphocyte-restricted expression to a reporter construct. Our work and that of others subsequently revealed that RAG-2 promoter expression is positively regulated by BSAP (PAX-5) and c-Myb transcription factors in B- and T-lineage cells, respectively. Although BSAP and c-Myb were deemed necessary for lymphocyte-specific expression, our analysis also uncovered a G-rich region at the 5′-end of the core promoter that was essential for full activity in lymphocyte cell lines. Site-directed mutagenesis revealed that a GA-box within the G-rich region was required for full promoter activity and subsequent DNA binding assays demonstrated that this element was specifically recognized by Sp1. Apart from showing that Sp1 interacts within the RAG-2 promoter, we also demonstrate that the Sp1-binding site is necessary for the high-level activation of this promoter.",
author = "Miranda, {Gustavo A.} and Maria Villalvazo and Zoran Galic and Jackelyn Alva and Roxanna Abrines and Yvette Yates and Evans, {Cory J.} and Aguilera, {Renato J.}",
year = "2002",
month = "6",
day = "19",
doi = "10.1016/S0161-5890(02)00007-X",
language = "English (US)",
volume = "38",
pages = "1151--1159",
journal = "Molecular Immunology",
issn = "0161-5890",
publisher = "Elsevier Limited",
number = "15",

}

TY - JOUR

T1 - Combinatorial regulation of the murine RAG-2 promoter by Sp1 and distinct lymphocyte-specific transcription factors

AU - Miranda, Gustavo A.

AU - Villalvazo, Maria

AU - Galic, Zoran

AU - Alva, Jackelyn

AU - Abrines, Roxanna

AU - Yates, Yvette

AU - Evans, Cory J.

AU - Aguilera, Renato J.

PY - 2002/6/19

Y1 - 2002/6/19

N2 - The recombination activation genes, RAG-1 and RAG-2, encode the critical components of the recombinase complex responsible for the generation of functional antigen receptor genes. In order to gain an insight into the transcription factors and cis-acting elements that regulate the lymphocyte-specific expression of RAG-2, the promoter-region of this gene was isolated and characterized. This analysis demonstrated that a relatively small promoter fragment could confer lymphocyte-restricted expression to a reporter construct. Our work and that of others subsequently revealed that RAG-2 promoter expression is positively regulated by BSAP (PAX-5) and c-Myb transcription factors in B- and T-lineage cells, respectively. Although BSAP and c-Myb were deemed necessary for lymphocyte-specific expression, our analysis also uncovered a G-rich region at the 5′-end of the core promoter that was essential for full activity in lymphocyte cell lines. Site-directed mutagenesis revealed that a GA-box within the G-rich region was required for full promoter activity and subsequent DNA binding assays demonstrated that this element was specifically recognized by Sp1. Apart from showing that Sp1 interacts within the RAG-2 promoter, we also demonstrate that the Sp1-binding site is necessary for the high-level activation of this promoter.

AB - The recombination activation genes, RAG-1 and RAG-2, encode the critical components of the recombinase complex responsible for the generation of functional antigen receptor genes. In order to gain an insight into the transcription factors and cis-acting elements that regulate the lymphocyte-specific expression of RAG-2, the promoter-region of this gene was isolated and characterized. This analysis demonstrated that a relatively small promoter fragment could confer lymphocyte-restricted expression to a reporter construct. Our work and that of others subsequently revealed that RAG-2 promoter expression is positively regulated by BSAP (PAX-5) and c-Myb transcription factors in B- and T-lineage cells, respectively. Although BSAP and c-Myb were deemed necessary for lymphocyte-specific expression, our analysis also uncovered a G-rich region at the 5′-end of the core promoter that was essential for full activity in lymphocyte cell lines. Site-directed mutagenesis revealed that a GA-box within the G-rich region was required for full promoter activity and subsequent DNA binding assays demonstrated that this element was specifically recognized by Sp1. Apart from showing that Sp1 interacts within the RAG-2 promoter, we also demonstrate that the Sp1-binding site is necessary for the high-level activation of this promoter.

UR - http://www.scopus.com/inward/record.url?scp=0036278503&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036278503&partnerID=8YFLogxK

U2 - 10.1016/S0161-5890(02)00007-X

DO - 10.1016/S0161-5890(02)00007-X

M3 - Article

C2 - 12044781

AN - SCOPUS:0036278503

VL - 38

SP - 1151

EP - 1159

JO - Molecular Immunology

JF - Molecular Immunology

SN - 0161-5890

IS - 15

ER -