Transcriptional regulation of mouse αB- and γF-crystallin genes in lens

Opposite promoter-specific interactions between Pax6 and large Maf transcription factors

Ying Yang, Bharesh K. Chauhan, Kveta Cveklova, Ales Cvekl

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Mammalian αB-crystallin is highly expressed both in lens epithelium and lens fibers. In contrast, γF-crystallin is highly expressed in the lens fiber cells. Crystallin gene expression in lens is regulated at the level of transcription by a sparse number of specific DNA-binding transcription factors. Here, we report studies on transcriptional regulation of mouse αB- and γF-crystallin promoters by specific combinations of Pax6/Pax6(5a), large Mafs (MafA, MafB, c-Maf, and NRL), Sox1, Sox2, Six3, and RARβ/RXRβ. Two sets of these factors, co-expressed both in lens epithelium and in lens fibers, were tested in co-transfection assays using cultured lens and non-lens cells. Regulation of αB-crystallin was studied in the presence of lens epithelial-factors Pax6, MafB, and RARβ/RXRβ, and lens fiber-factors Pax6, MafA, c-Maf, and NRL. Pax6 proteins activated the αB-crystallin promoter (-162 to +45) with any combination of Mafs. Addition of RARβ/RXRβ further increased its promoter activity. Gel shift assays using lens nuclear extracts demonstrated interactions of Pax6, Maf, and retinoic acid nuclear receptor proteins with two lens-specific regions, the distal LSR1 (-147/-118) and proximal LSR2 (-78/-40), of the αB-crystallin promoter. In contrast, Pax6 proteins acted as repressors of γF-crystallin promoter activity elicited by a combination of large Mafs, Sox, and RARβ/RXRβ proteins in transiently transfected lens and non-lens cells. The results show that Pax6 conversely regulates these two lens crystallin promoters. We propose that the opposite roles of Pax6 in crystallin gene regulation are results of different promoter architectures of the αB- and γF-crystallin genes, developmentally regulated association of transcription factors with the corresponding cis-regulatory sites, and specific recruitment of transcriptional co-activators and co-repressors by Pax6.

Original languageEnglish (US)
Pages (from-to)351-368
Number of pages18
JournalJournal of Molecular Biology
Volume344
Issue number2
DOIs
StatePublished - Nov 19 2004

Fingerprint

Large Maf Transcription Factors
Crystallins
Lenses
Genes
Transcription Factors
Epithelium

Keywords

  • crystallins
  • lens
  • Maf
  • Pax6
  • Six3, Sox1, Sox2

ASJC Scopus subject areas

  • Virology

Cite this

Transcriptional regulation of mouse αB- and γF-crystallin genes in lens : Opposite promoter-specific interactions between Pax6 and large Maf transcription factors. / Yang, Ying; Chauhan, Bharesh K.; Cveklova, Kveta; Cvekl, Ales.

In: Journal of Molecular Biology, Vol. 344, No. 2, 19.11.2004, p. 351-368.

Research output: Contribution to journalArticle

@article{be3ab85532a44826bd694a726b189841,
title = "Transcriptional regulation of mouse αB- and γF-crystallin genes in lens: Opposite promoter-specific interactions between Pax6 and large Maf transcription factors",
abstract = "Mammalian αB-crystallin is highly expressed both in lens epithelium and lens fibers. In contrast, γF-crystallin is highly expressed in the lens fiber cells. Crystallin gene expression in lens is regulated at the level of transcription by a sparse number of specific DNA-binding transcription factors. Here, we report studies on transcriptional regulation of mouse αB- and γF-crystallin promoters by specific combinations of Pax6/Pax6(5a), large Mafs (MafA, MafB, c-Maf, and NRL), Sox1, Sox2, Six3, and RARβ/RXRβ. Two sets of these factors, co-expressed both in lens epithelium and in lens fibers, were tested in co-transfection assays using cultured lens and non-lens cells. Regulation of αB-crystallin was studied in the presence of lens epithelial-factors Pax6, MafB, and RARβ/RXRβ, and lens fiber-factors Pax6, MafA, c-Maf, and NRL. Pax6 proteins activated the αB-crystallin promoter (-162 to +45) with any combination of Mafs. Addition of RARβ/RXRβ further increased its promoter activity. Gel shift assays using lens nuclear extracts demonstrated interactions of Pax6, Maf, and retinoic acid nuclear receptor proteins with two lens-specific regions, the distal LSR1 (-147/-118) and proximal LSR2 (-78/-40), of the αB-crystallin promoter. In contrast, Pax6 proteins acted as repressors of γF-crystallin promoter activity elicited by a combination of large Mafs, Sox, and RARβ/RXRβ proteins in transiently transfected lens and non-lens cells. The results show that Pax6 conversely regulates these two lens crystallin promoters. We propose that the opposite roles of Pax6 in crystallin gene regulation are results of different promoter architectures of the αB- and γF-crystallin genes, developmentally regulated association of transcription factors with the corresponding cis-regulatory sites, and specific recruitment of transcriptional co-activators and co-repressors by Pax6.",
keywords = "crystallins, lens, Maf, Pax6, Six3, Sox1, Sox2",
author = "Ying Yang and Chauhan, {Bharesh K.} and Kveta Cveklova and Ales Cvekl",
year = "2004",
month = "11",
day = "19",
doi = "10.1016/j.jmb.2004.07.102",
language = "English (US)",
volume = "344",
pages = "351--368",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Transcriptional regulation of mouse αB- and γF-crystallin genes in lens

T2 - Opposite promoter-specific interactions between Pax6 and large Maf transcription factors

AU - Yang, Ying

AU - Chauhan, Bharesh K.

AU - Cveklova, Kveta

AU - Cvekl, Ales

PY - 2004/11/19

Y1 - 2004/11/19

N2 - Mammalian αB-crystallin is highly expressed both in lens epithelium and lens fibers. In contrast, γF-crystallin is highly expressed in the lens fiber cells. Crystallin gene expression in lens is regulated at the level of transcription by a sparse number of specific DNA-binding transcription factors. Here, we report studies on transcriptional regulation of mouse αB- and γF-crystallin promoters by specific combinations of Pax6/Pax6(5a), large Mafs (MafA, MafB, c-Maf, and NRL), Sox1, Sox2, Six3, and RARβ/RXRβ. Two sets of these factors, co-expressed both in lens epithelium and in lens fibers, were tested in co-transfection assays using cultured lens and non-lens cells. Regulation of αB-crystallin was studied in the presence of lens epithelial-factors Pax6, MafB, and RARβ/RXRβ, and lens fiber-factors Pax6, MafA, c-Maf, and NRL. Pax6 proteins activated the αB-crystallin promoter (-162 to +45) with any combination of Mafs. Addition of RARβ/RXRβ further increased its promoter activity. Gel shift assays using lens nuclear extracts demonstrated interactions of Pax6, Maf, and retinoic acid nuclear receptor proteins with two lens-specific regions, the distal LSR1 (-147/-118) and proximal LSR2 (-78/-40), of the αB-crystallin promoter. In contrast, Pax6 proteins acted as repressors of γF-crystallin promoter activity elicited by a combination of large Mafs, Sox, and RARβ/RXRβ proteins in transiently transfected lens and non-lens cells. The results show that Pax6 conversely regulates these two lens crystallin promoters. We propose that the opposite roles of Pax6 in crystallin gene regulation are results of different promoter architectures of the αB- and γF-crystallin genes, developmentally regulated association of transcription factors with the corresponding cis-regulatory sites, and specific recruitment of transcriptional co-activators and co-repressors by Pax6.

AB - Mammalian αB-crystallin is highly expressed both in lens epithelium and lens fibers. In contrast, γF-crystallin is highly expressed in the lens fiber cells. Crystallin gene expression in lens is regulated at the level of transcription by a sparse number of specific DNA-binding transcription factors. Here, we report studies on transcriptional regulation of mouse αB- and γF-crystallin promoters by specific combinations of Pax6/Pax6(5a), large Mafs (MafA, MafB, c-Maf, and NRL), Sox1, Sox2, Six3, and RARβ/RXRβ. Two sets of these factors, co-expressed both in lens epithelium and in lens fibers, were tested in co-transfection assays using cultured lens and non-lens cells. Regulation of αB-crystallin was studied in the presence of lens epithelial-factors Pax6, MafB, and RARβ/RXRβ, and lens fiber-factors Pax6, MafA, c-Maf, and NRL. Pax6 proteins activated the αB-crystallin promoter (-162 to +45) with any combination of Mafs. Addition of RARβ/RXRβ further increased its promoter activity. Gel shift assays using lens nuclear extracts demonstrated interactions of Pax6, Maf, and retinoic acid nuclear receptor proteins with two lens-specific regions, the distal LSR1 (-147/-118) and proximal LSR2 (-78/-40), of the αB-crystallin promoter. In contrast, Pax6 proteins acted as repressors of γF-crystallin promoter activity elicited by a combination of large Mafs, Sox, and RARβ/RXRβ proteins in transiently transfected lens and non-lens cells. The results show that Pax6 conversely regulates these two lens crystallin promoters. We propose that the opposite roles of Pax6 in crystallin gene regulation are results of different promoter architectures of the αB- and γF-crystallin genes, developmentally regulated association of transcription factors with the corresponding cis-regulatory sites, and specific recruitment of transcriptional co-activators and co-repressors by Pax6.

KW - crystallins

KW - lens

KW - Maf

KW - Pax6

KW - Six3, Sox1, Sox2

UR - http://www.scopus.com/inward/record.url?scp=7444233073&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=7444233073&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2004.07.102

DO - 10.1016/j.jmb.2004.07.102

M3 - Article

VL - 344

SP - 351

EP - 368

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 2

ER -