TY - JOUR
T1 - Dual roles for proxl in the regulation of the chicken β3B1-crystallin promoter
AU - Chen, Xiaoren
AU - Taube, Jennifer R.
AU - Simirskii, Vladimir I.
AU - Patel, Tapan P.
AU - Duncan, Melinda K.
PY - 2008/4
Y1 - 2008/4
N2 - 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.
AB - 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.
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U2 - 10.1167/iovs.07-1300
DO - 10.1167/iovs.07-1300
M3 - Article
C2 - 18385074
AN - SCOPUS:45549108010
SN - 0146-0404
VL - 49
SP - 1542
EP - 1552
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
IS - 4
ER -