The Na-G ion channel is transcribed from a single promoter controlled by distinct neuron- and Schwann cell-specific DNA elements.
Na-G is a putative sodium (or cationic) channel expressed in neurons and glia of the PNS, in restricted neuronal subpopulations of the brain, and in several tissues outside the nervous system, like lung and adrenal medulla. To analyze the mechanisms underlying tissue-specific expression of this channel, we isolated the 5' region of the corresponding gene and show that Na-G mRNA transcription proceeds from a single promoter with multiple initiation sites. By transgenic mice studies, we demonstrate that 600 bp containing the Na-G proximal promoter region and the first exon are sufficient to drive the expression of a beta-galactosidase reporter gene in neurons of both CNS and PNS, whereas expression in Schwann cells depends on more remote DNA elements lying in the region between -6,500 and -1,050 bp upstream of the main transcription initiation sites. Crucial elements for lung-specific expression seem to be located in the region between -1,050 and -375 bp upstream of the promoter. Using in vivo footprint experiments, we demonstrate that several sites of the Na-G proximal promoter region are bound specifically by nuclear proteins in dorsal root ganglion neurons, as compared with nonexpressing hepatoma cells.
Poiraud
E
E
Biochimie Cellulaire, CNRS UPR 9065, Collège de France, Paris.
Gruszczynski
C
C
Porteu
A
A
Cambier
H
H
Escurat
M
M
Koulakoff
A
A
Kahn
A
A
Berwald-Netter
Y
Y
Gautron
S
S
D000818:Animals; D001483:Base Sequence; D002490:Central Nervous System / Q000378:metabolism; D018983:DNA Footprinting; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons / Q000235:genetics; D005727:Ganglia, Spinal / Q000378:metabolism; D017930:Genes, Reporter; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics*; D009474:Neurons / Q000378:metabolism*; D009475:Neurons, Afferent / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D009928:Organ Specificity; D017933:Peripheral Nervous System / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D011916:Rats, Inbred F344; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D012583:Schwann Cells / Q000378:metabolism*; D015222:Sodium Channels / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D061566:Voltage-Gated Sodium Channels; D001616:beta-Galactosidase / Q000096:biosynthesis
D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C402116:Scn7a protein, rat; D015222:Sodium Channels; D061566:Voltage-Gated Sodium Channels; D001616:beta-Galactosidase