VISUALIZATION OF CENTROMERE PROTEINS CENP-B AND CENP-C ON A STABLE DICENTRIC CHROMOSOME IN CYTOLOGICAL SPREADS
VISUALIZATION OF CENTROMERE PROTEINS CENP-B AND CENP-C ON A STABLE DICENTRIC CHROMOSOME IN CYTOLOGICAL SPREADS
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DOI：
10.1007/bf00293329
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发表时间：
1989-06-01
期刊：
影响因子：
1.6
通讯作者：
STETTEN, G
中科院分区：
文献类型：
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作者：
EARNSHAW, WC;RATRIE, H;STETTEN, G
我们已经筛选了两个着丝粒自身抗原，CENP-B（80 kDa）和CENP-C（140 kDa）的存在下，在一个自然发生的稳定的双着丝粒染色体的非活性着丝粒使用特异性抗体，不与任何其他染色体蛋白交叉反应。为了区分该染色体上的活性和非活性着丝粒，我们已经开发了一种标准甲醇/乙酸固定程序的修改，使我们能够获得高质量的细胞学涂片，保留抗着丝粒抗体的抗原性。我们已经注意到特异性抗CENP-B和CENP-C抗体的免疫染色模式的三个差异。(1)可检测的CENP-B的量因染色体而异。CENP-C的量在所有染色体上似乎或多或少相同。(2)CENP-B存在于稳定的双着丝粒常染色体的活性和非活性着丝粒处。CENP-C在非活性着丝粒处不可检测。(3)虽然用抗CENP-C抗体的免疫荧光通常给出两个离散的斑点，但用抗CENP-B的染色通常表现为连接两个姐妹着丝粒的单个亮条。这表明CENP-C可能局限于着丝粒区域的外着丝粒，而CENP-B可能分布在整个着丝粒中。我们的数据表明，CENP-C可能是一些不变的染色体亚结构，如动粒的组成部分。CENP-B还可能参与着丝粒的其他功能，如染色体的移动和DNA的包装。
We have screened for the presence of two centromere autoantigens, CENP-B (80 kDa) and CENP-C (140 kDa) at the inactive centromere of a naturally occuring stable dicentric chromosome using specific antibodies that do not cross-react with any other chromsomal proteins. In order to discriminant between the active and inactive centromeres on this chromosome we have developed a modification of the standard methanol/acetic acid fixation procedure that allows us to obtain high-quality cytological spreads that retain antigenicity with the anti-centromere antibodies. We have noted three differences in the immunostaining patterns with specific anti-CENP-B and CENP-C antibodies. (1) The amount of detectable CENP-B varies from chromosome to chromosome. The amount of CENP-C appears to be more or less the same on all chromosomes. (2) CENP-B is present at both active and inactive centromeres of stable dicentric autosomes. CENP-C is not detectable at the inactive centromeres. (3) While immmunofluorscence with anti-CENP-C antibodies typically gives two discrete spots, staining with anti-CENP-B often appears as a single bright bar connecting both sister centromeres. This suggests that while CENP-C may be confined to the outer centromere in the kinetochore region, CENP-B may be distributed throughout the entire centromere. Our data suggest that CENP-C is likely to be a component of some invariant chromosomal substructure, such as the kinetochore. CENP-B may be involved in some other apsect of centromere function, such as chromosome movement or DNA packaging.