Analysis of non-crystallin lens fiber cell gene expression in c-Maf -/- mice

Daryle J. DePianto, Tom N. Blankenship, John F. Hess, Paul G FitzGerald

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Purpose: To examine the role of the c-Maf transcription factor in the regulation of non-crystallin lens fiber cell specific gene expression. Methods: Expression of lens fiber cell genes in c-Maf null mice was analyzed by immunohistochemical and RT-PCR methods. In addition, the effect of c-Maf on fiber cell gene promoter fragments was examined in co-transfection experiments. Results: Examination of protein expression in c-Maf homozygous mutant mice revealed undetectable levels of CP49, CP115 and MIP protein. mRNA transcripts for these non-crystallin genes were observed at reduced levels in subsequent RT-PCR experiments. The co-transfection of a c-Maf expression construct with reporter constructs containing the proximal promoters of the CP49 and CP115 genes, the structural components of the lens fiber cell beaded filament, did not result in activation of reporter gene expression, as was observed with a crystallin promoter. Conclusions: While c-Maf has been shown to play a critical role in lens specific expression of the numerous crystallin genes, it does not appear to regulate some non-crystallin fiber cell genes in the same manner. In contrast to most crystallin genes, detectable levels of CP49, CP115, and MIP transcripts are achieved in the absence of c-Maf. In co-transfection experiments, the CP49 and CP115 proximal promoter sequences were not activated by c-Maf. This suggests that increases in transcription of non-crystallin lens fiber cell genes can be enacted in a c-Maf independent fashion.

Original languageEnglish (US)
Pages (from-to)288-294
Number of pages7
JournalMolecular Vision
Volume9
StatePublished - 2003

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Lenses
Gene Expression
Crystallins
Genes
Transfection
Maf Transcription Factors
Gene Components
Polymerase Chain Reaction
Reporter Genes
Proteins
Messenger RNA

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Analysis of non-crystallin lens fiber cell gene expression in c-Maf -/- mice. / DePianto, Daryle J.; Blankenship, Tom N.; Hess, John F.; FitzGerald, Paul G.

In: Molecular Vision, Vol. 9, 2003, p. 288-294.

Research output: Contribution to journalArticle

DePianto, Daryle J. ; Blankenship, Tom N. ; Hess, John F. ; FitzGerald, Paul G. / Analysis of non-crystallin lens fiber cell gene expression in c-Maf -/- mice. In: Molecular Vision. 2003 ; Vol. 9. pp. 288-294.
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N2 - Purpose: To examine the role of the c-Maf transcription factor in the regulation of non-crystallin lens fiber cell specific gene expression. Methods: Expression of lens fiber cell genes in c-Maf null mice was analyzed by immunohistochemical and RT-PCR methods. In addition, the effect of c-Maf on fiber cell gene promoter fragments was examined in co-transfection experiments. Results: Examination of protein expression in c-Maf homozygous mutant mice revealed undetectable levels of CP49, CP115 and MIP protein. mRNA transcripts for these non-crystallin genes were observed at reduced levels in subsequent RT-PCR experiments. The co-transfection of a c-Maf expression construct with reporter constructs containing the proximal promoters of the CP49 and CP115 genes, the structural components of the lens fiber cell beaded filament, did not result in activation of reporter gene expression, as was observed with a crystallin promoter. Conclusions: While c-Maf has been shown to play a critical role in lens specific expression of the numerous crystallin genes, it does not appear to regulate some non-crystallin fiber cell genes in the same manner. In contrast to most crystallin genes, detectable levels of CP49, CP115, and MIP transcripts are achieved in the absence of c-Maf. In co-transfection experiments, the CP49 and CP115 proximal promoter sequences were not activated by c-Maf. This suggests that increases in transcription of non-crystallin lens fiber cell genes can be enacted in a c-Maf independent fashion.

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