Abstract
Differentiated retinal pigment epithelial (RPE) cells in vive express basal levels of FGF-5, a secreted member of the FGF gene family. RPE cells proliferate in response to pathological events, resulting in a transient increase in FGF-5 gene expression. The goal of this study is to identify cis- acting sequences in the FGF-5 gene promoter which upregulate FGF-5 gene expression when differentiated RPE cells enter the cell cycle and proliferate. In vitro cultures of RPE cells were transfected with various FGF-5 promoter/luciferase deletion constructs, using methods specifically optimized for proliferating and differentiated RPE cells. A proximal promoter/enhancer whose activity is not cell-context dependent was identified between FGF-5 sequences - 314 and +48. In addition, a silencer element (- 1256/-883) was identified in the distal region which is active only in differentiated RPE cells. When tested in a heterologous system, the same element had silencer activity in differentiated cells. Two small regions in the distal FGF-5 gene promoter, -1195/-1173 and -984/-967 were able to specifically bind to nuclear proteins from differentiated RPE cells but not from proliferating RPE cells as evidenced by gel mobility shift assays. Therefore, FGF-5 gene expression in the RPE may be regulated by the formation of differentiation-specific complexes.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 151-162 |
| Number of pages | 12 |
| Journal | Experimental Eye Research |
| Volume | 67 |
| Issue number | 2 |
| DOIs | |
| State | Published - Aug 1998 |
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Keywords
- Differentiation
- Fibroblast growth factors
- Retinal pigment epithelium
- Transcription
ASJC Scopus subject areas
- Ophthalmology
- Sensory Systems
Cite this
Differentiation of retinal pigment epithelial cells in vitro uncovers silencer activity in the FGF-5 gene promoter. / Gelfman, Claire Mazow; Kelleher, Cassandra M.; Hjelmeland, Leonard M.
In: Experimental Eye Research, Vol. 67, No. 2, 08.1998, p. 151-162.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Differentiation of retinal pigment epithelial cells in vitro uncovers silencer activity in the FGF-5 gene promoter
AU - Gelfman, Claire Mazow
AU - Kelleher, Cassandra M.
AU - Hjelmeland, Leonard M
PY - 1998/8
Y1 - 1998/8
N2 - Differentiated retinal pigment epithelial (RPE) cells in vive express basal levels of FGF-5, a secreted member of the FGF gene family. RPE cells proliferate in response to pathological events, resulting in a transient increase in FGF-5 gene expression. The goal of this study is to identify cis- acting sequences in the FGF-5 gene promoter which upregulate FGF-5 gene expression when differentiated RPE cells enter the cell cycle and proliferate. In vitro cultures of RPE cells were transfected with various FGF-5 promoter/luciferase deletion constructs, using methods specifically optimized for proliferating and differentiated RPE cells. A proximal promoter/enhancer whose activity is not cell-context dependent was identified between FGF-5 sequences - 314 and +48. In addition, a silencer element (- 1256/-883) was identified in the distal region which is active only in differentiated RPE cells. When tested in a heterologous system, the same element had silencer activity in differentiated cells. Two small regions in the distal FGF-5 gene promoter, -1195/-1173 and -984/-967 were able to specifically bind to nuclear proteins from differentiated RPE cells but not from proliferating RPE cells as evidenced by gel mobility shift assays. Therefore, FGF-5 gene expression in the RPE may be regulated by the formation of differentiation-specific complexes.
AB - Differentiated retinal pigment epithelial (RPE) cells in vive express basal levels of FGF-5, a secreted member of the FGF gene family. RPE cells proliferate in response to pathological events, resulting in a transient increase in FGF-5 gene expression. The goal of this study is to identify cis- acting sequences in the FGF-5 gene promoter which upregulate FGF-5 gene expression when differentiated RPE cells enter the cell cycle and proliferate. In vitro cultures of RPE cells were transfected with various FGF-5 promoter/luciferase deletion constructs, using methods specifically optimized for proliferating and differentiated RPE cells. A proximal promoter/enhancer whose activity is not cell-context dependent was identified between FGF-5 sequences - 314 and +48. In addition, a silencer element (- 1256/-883) was identified in the distal region which is active only in differentiated RPE cells. When tested in a heterologous system, the same element had silencer activity in differentiated cells. Two small regions in the distal FGF-5 gene promoter, -1195/-1173 and -984/-967 were able to specifically bind to nuclear proteins from differentiated RPE cells but not from proliferating RPE cells as evidenced by gel mobility shift assays. Therefore, FGF-5 gene expression in the RPE may be regulated by the formation of differentiation-specific complexes.
KW - Differentiation
KW - Fibroblast growth factors
KW - Retinal pigment epithelium
KW - Transcription
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UR - http://www.scopus.com/inward/citedby.url?scp=0032144575&partnerID=8YFLogxK
U2 - 10.1006/exer.1998.0506
DO - 10.1006/exer.1998.0506
M3 - Article
C2 - 9733582
AN - SCOPUS:0032144575
VL - 67
SP - 151
EP - 162
JO - Experimental Eye Research
JF - Experimental Eye Research
SN - 0014-4835
IS - 2
ER -