Mouse 3-phosphoglycerate dehydrogenase gene: genomic organization, chromosomal localization, and promoter analysis.
Mouse 3-phosphoglycerate dehydrogenase gene: genomic organization, chromosomal localization, and promoter analysis.
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DOI：
10.1016/j.gene.2004.03.015
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发表时间：
2004-06
期刊：
影响因子：
3.5
通讯作者：
J. Mitoma;S. Furuya;M. Shimizu;Y. Shinoda;Kazuyuki Yoshida;N. Azuma;Hideyuki Tanaka;Yasuo Suzuki;Y. Hirabayashi
中科院分区：
文献类型：
--
作者：
J. Mitoma;S. Furuya;M. Shimizu;Y. Shinoda;Kazuyuki Yoshida;N. Azuma;Hideyuki Tanaka;Yasuo Suzuki;Y. Hirabayashi
d-3-磷酸甘油酸脱氢酶（Phgdh; EC 1.1.1.95）是磷酸化途径中L-丝氨酸生物合成的第一个关键酶。我们最近已经证明，在发育和成熟的脑中，Phgdh的表达以细胞谱系特异性方式高度调节，主要在神经上皮干细胞、放射状神经胶质细胞和星形胶质细胞中（J. Neurosci. 21（2001）7691; Arch.Histol.Cytol.66（2003）109）。为了深入了解Phgdh表达的调控机制，我们分离了一个包含整个小鼠Phgdh基因的小鼠基因组克隆。结构分析表明，Phgdh基因跨越约27个碱基（kb）的长度和包含12个外显子与11个插入内含子。使用荧光原位杂交（FISH），我们将该基因定位到小鼠3号染色体F2-F3区。对5′侧翼区的1.8kb片段的分析表明，在转录起始位点附近的经典TATA盒基序缺失。相反，富含GC的近端区域含有潜在的Sp1识别序列，该区域在小鼠，大鼠和人类中是保守的。瞬时转染分析表明，Phgdh基础转录所需的顺式作用元件包含在启动子的−196/+4近端序列中，其中保守的Sp1识别位点对基础启动子活性起重要作用。
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.