The mouse p97 (CDC48) gene -: Genomic structure, definition of transcriptional regulatory sequences, gene expression, and characterization of a pseudogene
The mouse p97 (CDC48) gene -: Genomic structure, definition of transcriptional regulatory sequences, gene expression, and characterization of a pseudogene
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DOI：
10.1074/jbc.274.15.10154
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发表时间：
1999-04-09
影响因子：
4.8
通讯作者：
Shima, DT
中科院分区：
文献类型：
--
作者：
Müller, JMM;Meyer, HH;Shima, DT
在这里，我们首次描述了小鼠 p97(CDC48) AAA ATP 酶的基因组组织、转录调控序列以及成体和胚胎基因表达。在基因组文库筛选中分离出代表两个不同 p97 基因的克隆，其中一个可能代表非功能性加工假基因。编码功能性 mRNA 的基因的编码区被 16 个内含子中断，包含 20.4 千碱基对。转录起始位点的定义和序列分析表明，该基因包含一个无 TATA、富含 CC 的启动子区域，其起始元件跨越转录起始位点。通过瞬时转染测定确定，基础转录活性所需的顺式作用元件位于侧翼区域的 410 个碱基对内。在免疫组织学分析中，p97 在胚胎和成人中广泛表达，但蛋白质水平以细胞类型和细胞分化依赖性方式受到严格控制。在给定组织内的细胞水平上发现 p97 免疫染色显着的异质性，细胞质和细胞核中的蛋白质含量差异很大，表明 p97 具有高度调节和间歇性的功能。这项研究为详细分析 p97 的复杂调节以及使用小鼠靶向基因操作评估其功能意义所需的试剂提供了基础。
Here we present the first description of the genomic organization, transcriptional regulatory sequences, and adult and embryonic gene expression for the mouse p97(CDC48) AAA ATPase. Clones representing two distinct p97 genes were isolated in a genomic library screen, one of them likely representing a non-functional processed pseudogene. The coding region of the gene encoding the functional mRNA is interrupted by 16 introns and encompasses 20.4 kilobase pairs. Definition of the transcriptional initiation site and sequence analysis showed that the gene contains a TATA-less, CC-rich promoter region with an initiator element spanning the transcription start site. Cis-acting elements necessary for basal transcription activity reside within 410 base pairs of the flanking region as determined by transient transfection assays. In immunohistological analyses, p97 was widely expressed in embryos and adults, but protein levels were tightly controlled in a cell type- and cell differentiation-dependent manner. A remarkable heterogeneity in p97 immunostaining was found on a cellular level within a given tissue, and protein amounts in the cytoplasm and nucleus varied widely, suggesting a highly regulated and intermittent function for p97, This study provides the basis for a detailed analysis of the complex regulation of p97 and the reagents required for assessing its functional significance using targeted gene manipulation in the mouse.