Mole-Bajer J, Bajer A S, Zinkowski R P, Balczon R D, Brinkley B R
Department of Biology, University of Oregon, Eugene 97403.
Proc Natl Acad Sci U S A. 1990 May;87(9):3599-603. doi: 10.1073/pnas.87.9.3599.
Human autoantibodies from a patient with scleroderma CREST (calcinosis, Raynaud phenomenon, esophageal dismotility, sclerodactyly, telangiectasia) were used to immunostain kinetochores on chromosomes in endosperm of the seed of the monocot Haemanthus katherinae Bak. Kinetochores of mitotic chromosomes and prekinetochores of interphase cells were specifically stained using conventional indirect immunofluorescence procedures as well as a nonfading immunogold-silver-enhanced technique and analyzed by fluorescence and video microscopy. In interphase, prekinetochores were either single or double structures depending on the stage of the cell cycle but became quadruple (two distinct stained dots on each chromatid) in mid-to-late prophase. In favorable preparations of prometaphase chromosomes, multiple subunits could be resolved within each sister kinetochore suggesting a compound organization. Western blot analysis demonstrated common epitopes in centromeric peptides of HeLa and Haemanthus cell extracts. Although the molecular mass of individual polypeptides differed in the two species, the presence of shared epitopes indicates striking conservation of centromere/kinetochore components throughout evolution.
来自一名患有硬皮病CREST（钙质沉着、雷诺现象、食管动力障碍、指端硬化、毛细血管扩张）患者的人类自身抗体，被用于对单子叶植物海氏补血草种子胚乳中染色体上的动粒进行免疫染色。使用传统的间接免疫荧光程序以及不褪色免疫金银增强技术，对有丝分裂染色体的动粒和间期细胞的前动粒进行特异性染色，并通过荧光和视频显微镜进行分析。在间期，前动粒根据细胞周期阶段为单结构或双结构，但在前期中期到后期变为四重结构（每个染色单体上有两个明显的染色点）。在有丝分裂中期染色体的良好制片中，每个姐妹动粒内可分辨出多个亚基，提示其复合结构。蛋白质印迹分析表明，海拉细胞和海氏补血草细胞提取物的着丝粒肽中存在共同表位。尽管两个物种中单个多肽的分子量不同，但共同表位的存在表明在整个进化过程中着丝粒/动粒成分具有显著的保守性。
