H. Skala et al., UPSTREAM ELEMENTS INVOLVED IN-VIVO IN ACTIVATION OF THE BRAIN-SPECIFIC RAT ALDOLASE-C GENE - ROLE OF BINDING-SITES FOR POU AND WINGED HELIXPROTEINS, The Journal of biological chemistry, 273(48), 1998, pp. 31806-31814
The rat aldolase C gene encodes a glycolytic enzyme strongly expressed
in adult brain. We previously reported that a 115-base pair (bp) prom
oter fragment was able to ensure the brain-specific expression of the
chloramphenicol acetyltransferase (CAT) reporter gene in transgenic mi
ce, but only at a low level (Thomas, M., Makeh, I., Briand, P., Kahn,
A, and Skala, H. (1993) fur. J. Biochem. 218, 143-151). Here we show t
hat in vivo activation of this promoter at a high level requires coope
ration between an upstream 0.6-kilobase pair (kb) fragment and far ups
tream sequences. In the 0.6-kb region, a 28-bp DNA element is shown to
include overlapping in vitro binding sites for POU domain regulatory
proteins and for the Winged Helix hepatocyte nuclear factor-3 beta fac
tor. An hepatocyte nuclear factor-3 beta-binding site previously descr
ibed in the short proximal promoter fragment is also shown to interact
in vitro with POU proteins, although with a lower affinity than the 2
8-bp motif. Additional binding sites for POU factors were detected in
the upstream 0.6-kb sequences. Progressive deletion in this region res
ulted in decreased expression levels of the transgenes in mice, sugges
ting synergistic interactions between these multiple POU-binding sites
. We propose that DNA elements characterized by a dual binding specifi
city for both POU domain and Winged Helix transcription factors could
play an essential role in the brain-specific expression of the aldolas
e C gene and other neuronal genes.