Functional analysis of mouse 3-phosphoglycerate dehydrogenase (Phgdh) gene promoter in developing brain

Motohiro Shimizu, Shigeki Furuya, Yoko Shinoda, Junya Mitoma, Tadashi Okamura, Ichiro Miyoshi, Noriyuki Kasai, Yoshio Hirabayashi, Yasuo Suzuki

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

D-3-Phosphoglycerate dehydrogenase (Phgdh; EC 1.1.1.95) is a necessary enzyme for de novo L-serine biosynthesis via the phosphorylated pathway. Targeted disruption of the mouse Phgdh gene has been shown to result in embryonic lethality, accompanied by severe abnormalities in brain development. Phgdh is expressed exclusively by neuroepithelium and radial glia in developing brain and later mainly by astrocytes. To elucidate the molecular mechanism that regulates such cell-type-specific expression of Phgdh in developing brain, an upstream 3.5-kilobase-pair (kbp) region of the gene harboring the promoter was characterized in primary cultures and transgenic mice. Analysis of Phgdh 5′-nested deletions in transfected cultures indicated that overall reporter luciferase levels were higher in glial cultures than those in neuronal cultures. Although basal promoter activity of the gene appeared to depend on an Spl binding sequence residing between -193 and -184 in both glial and neuronal cultures, an upstream 5′-flanking region between -1,794 and -1,095 contributed to up-regulation of Phgdh transcription in a glial-cell-specific manner. In the cerebral cortex of transgenic mouse embryos, the Phgdh promoter-LacZ transgene DNA containing -1,794/+4 promoter sequences directed β-galactosidase (β-Gal) expression mainly to Phgdh-positive neuroepithelium and radial glia. This glial preference diminished when β-Gal expression was driven solely by the upstream 0.2-kbp minimal promoter. However, glial preference of β-Gal expression was restored by placing the 700-base-pair 5′-DNA segment upstream of the minimal promoter. These observations suggest the presence of cis-acting elements that confer the cell type specificity of Phgdh transcription in the distal promoter region.

Original languageEnglish
Pages (from-to)623-632
Number of pages10
JournalJournal of Neuroscience Research
Volume76
Issue number5
DOIs
Publication statusPublished - Jun 1 2004

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Phosphoglycerate Dehydrogenase
Neuroglia
Brain
Genes
Transgenic Mice
Galactosidases
5' Flanking Region
DNA
Luciferases
Transgenes
Genetic Promoter Regions
Base Pairing
Astrocytes
Cerebral Cortex
Serine

All Science Journal Classification (ASJC) codes

  • Cellular and Molecular Neuroscience

Cite this

Functional analysis of mouse 3-phosphoglycerate dehydrogenase (Phgdh) gene promoter in developing brain. / Shimizu, Motohiro; Furuya, Shigeki; Shinoda, Yoko; Mitoma, Junya; Okamura, Tadashi; Miyoshi, Ichiro; Kasai, Noriyuki; Hirabayashi, Yoshio; Suzuki, Yasuo.

In: Journal of Neuroscience Research, Vol. 76, No. 5, 01.06.2004, p. 623-632.

Research output: Contribution to journalArticle

Shimizu, M, Furuya, S, Shinoda, Y, Mitoma, J, Okamura, T, Miyoshi, I, Kasai, N, Hirabayashi, Y & Suzuki, Y 2004, 'Functional analysis of mouse 3-phosphoglycerate dehydrogenase (Phgdh) gene promoter in developing brain', Journal of Neuroscience Research, vol. 76, no. 5, pp. 623-632. https://doi.org/10.1002/jnr.20102
Shimizu, Motohiro ; Furuya, Shigeki ; Shinoda, Yoko ; Mitoma, Junya ; Okamura, Tadashi ; Miyoshi, Ichiro ; Kasai, Noriyuki ; Hirabayashi, Yoshio ; Suzuki, Yasuo. / Functional analysis of mouse 3-phosphoglycerate dehydrogenase (Phgdh) gene promoter in developing brain. In: Journal of Neuroscience Research. 2004 ; Vol. 76, No. 5. pp. 623-632.
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