Cloning and characterization of the chick NMDA receptor subunit-1 gene.
The N-methyl-D-aspartate family of glutamate receptors (NMDARs) are tetrameric cation channels including NR1, NR2, and possibly NR3 subunits. The physiological properties of the receptor are directly related to the subunit composition of the oligomer. Whereas NR1 is essential for the formation of functional channels, NR2 and NR3 play a modulatory role. This work reports, for the first time, the cloning of a non-mammalian NR1 gene, including the 5'-regulatory region. The chick gene spans 31 kb of genomic DNA sequence composed of 22 exons interrupted by 21 introns. The exon/intron organization and the deduced amino acid sequence up to the end of exon 19 showed 85% homology to mammalian NR1 cloned genes. Significant differences from mammals were found at the C-terminal region which in the chick gene, includes a novel exon (exon 20) previously identified at the mRNA level in the chick retina. The basal promoter activity was shown to reside within the proximal 377 bp of 5'-regulatory region. The transcriptional activity of the 5'-flanking region of the chick NR1 gene was shown to be higher in neuronally-differentiated PC12 cells and in chick retinal neurons, than in non-differentiated PC12 cells and Müller glia. Comparison of the 5'-flanking region of chick NR1 gene with mammalian NR1 genes suggests that, in spite of significant differences in the nucleotide sequence, they share common DNA binding sites such as RE1, SP1, AP2, CREB, NFkappaB, and MEF2; therefore, some of the molecular mechanisms involved in transcriptional regulation of NR1 gene expression could be conserved among species.
Zarain-Herzberg
Angel
A
Departamento de Bioquímica, Facultad de Medicina, UNAM, México.
Lee-Rivera
Irene
I
Rodríguez
Gabriela
G
López-Colomé
Ana María
AM
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002645:Chickens / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004247:DNA / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics*; D005809:Genes, Regulator / Q000235:genetics*; D015698:Genomic Library; D006801:Humans; D007438:Introns / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism*; D016716:PC12 Cells; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012160:Retina / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D004268:DNA-Binding Proteins; C409466:NR1 NMDA receptor; D016194:Receptors, N-Methyl-D-Aspartate; D014157:Transcription Factors; D004247:DNA