Injection of anticentromere antibodies in interphase disrupts events required for chromosome movement at mitosis.
Injection of anticentromere antibodies in interphase disrupts events required for chromosome movement at mitosis.
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DOI：
10.1083/jcb.111.4.1519
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发表时间：
1990-10
期刊：
影响因子：
--
通讯作者：
Earnshaw WC
中科院分区：
文献类型：
--
作者：
Bernat RL;Borisy GG;Rothfield NF;Earnshaw WC
我们已经使用自身抗体来探测三种人类着丝粒蛋白在有丝分裂中的功能。这些抗体在免疫印迹中识别三种人多肽：CENP-A（17 kD）、CENP-B（80 kD）和CENP-C（140 kD）。纯化的抗着丝粒抗体（ACA-IgG）在间期引入组织培养细胞时会破坏有丝分裂。我们已经确定了抗体抑制的两个执行点。在有丝分裂前大于或等于3小时将抗体注射到细胞核中可阻止染色体在随后的有丝分裂中进行正常的前中期运动。在G2晚期期间在细胞核中注射的抗体导致细胞停滞在中期。令人惊讶的是，在前期开始之后引入的抗体不阻断有丝分裂。这些结果表明，CENP抗原参与两个基本的间期事件，所需的着丝粒在有丝分裂中的行动。这些可能包括S期结束时着丝粒组装与α卫星DNA的复制协调以及前期开始的动粒结构成熟。
We have used autoantibodies to probe the function of three human centromere proteins in mitosis. These antibodies recognize three human polypeptides in immunoblots: CENP-A (17 kD), CENP-B (80 kD), and CENP-C (140 kD). Purified anticentromere antibodies (ACA-IgG) disrupt mitosis when introduced into tissue culture cells during interphase. We have identified two execution points for antibody inhibition. Antibodies injected into the nucleus greater than or equal to 3 h before mitosis prevent the chromosomes from undergoing normal prometaphase movements in the subsequent mitosis. Antibodies injected in the nucleus during late G2 cause cells to arrest in metaphase. Surprisingly, antibodies introduced subsequent to the beginning of prophase do not block mitosis. These results suggest that the CENP antigens are involved in two essential interphase events that are required for centromere action in mitosis. These may include centromere assembly coordinate with the replication of alpha-satellite DNA at the end of S phase and the structural maturation of the kinetochore that begins at prophase.