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Regulatory mechanisms for the MHC class III genes

MHC III 类基因的调控机制

基本信息

批准号：
61480156
负责人：
TAKAHASHI Morinobu
金额：
$ 4.48万
依托单位：
Cancer Institute, Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

Mouse C4 and Sex-limited protein(Slp) are encoded by two recently duplicated genes of the major histocompatibility complex III of the mouse. The availability of many expression variants for these genes among mouse strains makes them the attractive system in which the regulation of eukaryotic genes under developmental, hormonal and tissue specific control can be studied. We isolated and characterized cDNA and genomic DNA clones for these genes from several mouse strains which show interesting variation for the expression of C4 and slp. Our experimental results obtained by various molecular genetic techniques can be summarized as follows.1. The mouse strains carrying H-2^k haplotype are characterized as low C4 producers. We measured C4 mRNA by Northern blotting and nuclear C4 RNA by nuclear run on assay using the hepatocytes from high C4 producing mouse(B10) and low C4 producing mouse(B10.BR). Furthermore we tested the 5'flanking regions of the C4 genes from these strains for their trans … More criptional regulatory activity with transfected cells. The results clearly showed that the low C4 bisynthesis is mainly controlled at the posttranscriptional and pretranslational level.2. To elucidate the molecular basis underlying the difference in the mode of expression of C4 gene(constitutional) and Slp gene(testosterone induced), we compared the nucleotide sequences of these two genes isolated from FM strain. Furthermore we tested the 5'flanking region fragments for transcriptional regulatory cativity using CAT assay in transfected cells. We identified the functional domains in the regions of these genes that appear to explain the difference in the gene expression of the C4 and Slp genes.3. We isolated and characterized all of C4-related genes from the cosmid library of C3H.W7 strain that shows an extraordinary mode of expression of Slp(constitutional expression). We found the three of apparent Slp genes indeed consisting of the C4-derived 5' half and Slp-derived 3' half. By comparing their nucleotide seguences with those of C4 and Slp genes, we concluded that these recombinant genes are derived by multiple cross over in the central portions of the gene. Less

小鼠C4和性别限制蛋白(SLP)是由小鼠主要组织相容性复合体III的两个最近复制的基因编码的。这些基因在小鼠品系中的许多表达变体使它们成为研究真核基因在发育、激素和组织特异性调控下的调控的诱人系统。我们从几个小鼠品系中分离并鉴定了这些基因的cDNA和基因组DNA克隆，这些克隆在C4和slp的表达上显示出有趣的差异。我们通过各种分子遗传学技术获得的实验结果可以概括如下。携带H-2^k单倍型的小鼠品系具有低C4产生的特点。用Northern印迹法检测高C4分泌小鼠(B10)和低C4分泌小鼠(B10.BR)的肝细胞C4mRNA和核Run-On实验。此外，我们还检测了这些菌株C4基因5‘侧翼区的反式…转基因细胞具有更强的功能调节活性。结果表明，低水平的C4双合成主要受转录后水平和翻译前水平的调控。为了阐明C4基因(组成基因)和SLP基因(睾酮诱导基因)表达模式差异的分子基础，我们比较了从FM株分离的这两个基因的核苷酸序列。此外，我们还利用CAT分析检测了5‘侧翼区片段在转基因细胞中的转录调控活性。我们确定了这些基因区域中的功能结构域，这些区域似乎解释了C4和SLP基因表达的差异。我们从C3H.W7株的粘粒文库中分离并鉴定了所有与C4相关的基因，该文库显示了SLP(结构性表达)的特殊表达模式。我们发现这三个SLP基因确实由C4来源的5‘端和SLP来源的3’端组成。通过比较它们与C4和SLP基因的核苷酸序列，我们得出结论，这些重组基因是通过基因中央部分的多次交叉而产生的。较少