Dual roles for proxl in the regulation of the chicken β3B1-crystallin promoter

Xiaoren Chen, Jennifer R. Taube, Vladimir I. Simirskii, Tapan Patel, Melinda K. Duncan

Research output: Contribution to journalArticle

Abstract

Purpose. Lens fiber cell differentiation is marked by the onset of β3B1-crystallin expression and is controlled by the cooperative action of a set of transcription factors including Prox1, an atypical homeodomain protein. Previously, the authors reported that Prox1 directly interacts with the OL2 element found in the chicken βB1-crystallin basal promoter to activate the expression of this gene. Here they mapped the location of activating and repressing sequences of the full-length chicken βB1-crystallin promoter (-432/+30) in lens epithelial cells, annular pad cells, and intact lens and characterized Prox1- binding sites found in this region. Methods. Transfection analysis and transgenic mice were used to characterize upstream regions of the chicken βB1-crystallin gene. DNaseI footprinting and chromatin immunoprecipitation was performed to identify Prox1-binding sites, and transfection analyses were used to characterize these sites functionally. Results. Sequences between - 152 and - 432 of the chicken βB1-crystallin promoter mediated either promoter activation or repression, depending on the stage of lens differentiation tested. Two new Prox1-binding sites were found in this region that bound Prox1 more avidly than the OL2 element. However, neither binding site conferred Prox1-mediated activation on a heterologous promoter; instead, each allowed Prox1 to repress promoter function. Conclusions. The function of the upstream region of the chicken βB1-crystallin promoter changes depending on cellular context. These data suggest that Prox1 function as a transcriptional activator could be regulated at the DNA level based on the characteristics of the responsive elements.

Original languageEnglish (US)
Pages (from-to)1542-1552
Number of pages11
JournalInvestigative Ophthalmology and Visual Science
Volume49
Issue number4
DOIs
StatePublished - Apr 1 2008
Externally publishedYes

Fingerprint

Crystallins
Chickens
Lenses
Binding Sites
Transfection
Homeodomain Proteins
Chromatin Immunoprecipitation
Transgenic Mice
Cell Differentiation
Transcription Factors
Epithelial Cells
Gene Expression
DNA
Genes

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Dual roles for proxl in the regulation of the chicken β3B1-crystallin promoter. / Chen, Xiaoren; Taube, Jennifer R.; Simirskii, Vladimir I.; Patel, Tapan; Duncan, Melinda K.

In: Investigative Ophthalmology and Visual Science, Vol. 49, No. 4, 01.04.2008, p. 1542-1552.

Research output: Contribution to journalArticle

Chen, Xiaoren ; Taube, Jennifer R. ; Simirskii, Vladimir I. ; Patel, Tapan ; Duncan, Melinda K. / Dual roles for proxl in the regulation of the chicken β3B1-crystallin promoter. In: Investigative Ophthalmology and Visual Science. 2008 ; Vol. 49, No. 4. pp. 1542-1552.
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