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首页> 外文期刊>Molecular and Cellular Biology >Stereospecific alignment of the X and Y elements is required for major histocompatibility complex class II DRA promoter function.
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Stereospecific alignment of the X and Y elements is required for major histocompatibility complex class II DRA promoter function.

机译：X和Y元素的立体定向比对是主要的组织相容性复杂的II类DRA启动子功能所必需的。

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摘要

The regulatory mechanisms controlling expression of the major histocompatibility complex (MHC) class II genes involve several cis-acting DNA elements, including the X and Y boxes. These two elements are conserved within all murine and human class II genes and are required for accurate and efficient transcription from MHC class II promoters. Interestingly, the distance between the X and Y elements is also evolutionarily conserved at 18 to 20 bp. To investigate the function of the invariant spacing in the human MHC class II gene, HLA-DRA, we constructed a series of spacing mutants which alters the distance between the X and Y elements by integral and half-integral turns of the DNA helix. Transient transfection of the spacing constructs into Raji cells revealed that inserting integral turns of the DNA helix (+20 and +10 bp) did not reduce promoter activity, while inserting or deleting half-integral turns of the DNA helix (+15, +5, and -5 bp) drastically reduced promoter activity. The loss of promoter function in these half-integral turn constructs was due neither to the inability of the X and Y elements to bind proteins nor to improper binding of the X- and Y-box-binding proteins. These data indicate that the X and Y elements must be aligned on the same side of the DNA helix to ensure normal function. This requirement for stereospecific alignment strongly suggests that the X- and Y-box-binding proteins either interact directly or are components of a larger transcription complex which assembles on one face of the DNA double helix.
机译：控制主要组织相容性复合体（MHC）II类基因表达的调节机制涉及几个顺式作用DNA元素，包括X和Y框。这两个元件在所有鼠类和人类II类基因中均是保守的，并且是从MHC II类启动子进行准确有效转录所需的。有趣的是，X和Y元素之间的距离在进化上也保守在18到20 bp。为了研究人类MHC II类基因HLA-DRA中不变间隔的功能，我们构建了一系列间隔突变体，该突变体通过DNA螺旋的整数和半整数匝改变X和Y元素之间的距离。间隔构建体的瞬时转染到Raji细胞中表明，插入DNA螺旋的整数匝（+20和+10 bp）不会降低启动子活性，而插入或删除DNA螺旋的半整数匝（+ 15，+ 5和-5 bp）大大降低了启动子活性。这些半整合转体构建体中启动子功能的丧失，既不是由于X和Y元素不能结合蛋白质，也不是由于X和Y盒结合蛋白的结合不当。这些数据表明X和Y元素必须在DNA螺旋的同一侧对齐，以确保正常功能。立体特异性比对的这种要求强烈表明，X-和Y-盒结合蛋白或者直接相互作用，或者是组装在DNA双螺旋的一个面上的较大转录复合物的组分。

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