Le. Lerner et al., Nrl and Sp nuclear proteins mediate transcription of rod-specific cGMP-phosphodiesterase beta-subunit gene - Involvement of multiple response elements, J BIOL CHEM, 276(37), 2001, pp. 34999-35007
cGMP-phosphodiesterase (PDE) is the key effector in rod photoreceptor signa
l transduction. Mutations in the gene encoding its catalytic, beta -subunit
(beta -PDE) cause retinal degenerations leading to blindness. We report th
at the short -93 to +53 sequence in the upstream region of this gene is suf
ficient for beta -PDE transcription in both Y79 human retinoblastoma cells
and Xenopus embryo heads maintained ex vivo. This sequence also functions a
s a minimal rod-specific promoter in transgenic Xenopus tadpoles. The Nrl t
ranscription factor binds in vitro to the beta Ap1/NRE regulatory element l
ocated within this region and transactivates it when overexpressed in non-r
etinal 293 embryonic kidney cells. We also found a G/C-rich activator eleme
nt, beta /GC, important for promoter activity in Y79 retinoblastoma cells a
nd Xenopus embryos. Both the ubiquitous Sp1 and the central nervous system-
specific Sp4 transcription factors are expressed in retina and interact wit
h this element in vitro. Electrophoretic mobilities of beta /GC-Y79 nuclear
protein complexes are altered by antibodies against Sp1 and Sp4. Thus, our
results implicate Nr1, Sp1, and Sp4 in transcriptional regulation of the r
od-specific minimal beta -PDE promoter. We also conclude that Xenopus laevi
s is an efficient system for analyzing the human beta -PDE promoter and may
be used to study other human retinal genes ex vivo and in vivo.