Mouse 3-phosphoglycerate dehydrogenase gene: genomic organization, chromosomal localization, and promoter analysis.
d-3-Phosphoglycerate dehydrogenase (Phgdh; EC 1.1.1.95) is the first committed enzyme of l-serine biosynthesis in the phosphorylated pathway. We have recently demonstrated that, in developing and mature brain, expression of Phgdh is highly regulated in a cell lineage-specific manner, mainly in neuroepithelial stem cells, radial glia, and astrocytes (J. Neurosci. 21 (2001) 7691; Arch. Histol. Cytol. 66 (2003) 109). To gain insight into the regulatory mechanism of Phgdh expression, we have isolated a mouse genomic clone that contains the entire mouse Phgdh gene. Structural analysis demonstrated that the Phgdh gene spans approximately 27 kilobases (kb) in length and comprises 12 exons with 11 intervening introns. Using fluorescent in situ hybridization (FISH), we mapped the gene to mouse chromosome 3, region F2-F3. Analysis of a 1.8 kb fragment of the 5'-flanking region showed that the classical TATA-box motif near transcription initiation sites was absent. Instead, a GC-rich proximal region containing a potential Sp1 recognition sequence was present; this region is conserved in mouse, rat, and human counterparts. Transient transfection analysis revealed that the cis-acting elements necessary for basal transcription of Phgdh are contained within the -196/+4 proximal sequence of the promoter, in which the conserved Sp1 recognition sites play an important role for basal promoter activity.
Mitoma
Junya
J
Glycobiology Program, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA.
Furuya
Shigeki
S
Shimizu
Motohiro
M
Shinoda
Yoko
Y
Yoshida
Kazuyuki
K
Azuma
Norihiro
N
Tanaka
Hideyuki
H
Suzuki
Yasuo
Y
Hirabayashi
Yoshio
Y
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D002237:Carbohydrate Dehydrogenases / Q000235:genetics*; D045744:Cell Line, Tumor; D002874:Chromosome Mapping; D033481:Chromosomes, Mammalian / Q000235:genetics; D005091:Exons; D005796:Genes / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D007438:Introns; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D050543:Phosphoglycerate Dehydrogenase; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D011993:Recombinant Fusion Proteins; D002237:Carbohydrate Dehydrogenases; D050543:Phosphoglycerate Dehydrogenase; D008156:Luciferases