Beenders Brent, Watrin Erwan, Legagneux Vincent, Kireev Igor, Bellini Michel
Department of Cell and Structural Biology, University of Illinois, Urbana, Illinois 61801, USA.
Chromosome Res. 2003;11(6):549-64. doi: 10.1023/a:1024999316867.
Several antibodies were used to examine the distribution of two condensin members, XCAP-E and XCAP-D2, in the nucleus of Xenopus oocytes. XCAP-D2 was found to be associated with the lampbrush chromosomes. The chromosomal regions containing XCAP-D2 correspond precisely to domains of highly compacted chromatin, suggesting a direct contribution of XCAP-D2 in meiotic chromatin organization. In contrast, XCAP-E was found to be absent from chromosomes but was detected at a high concentration in the granular component of nucleoli. The subnucleolar localization of XCAP-E was further confirmed by double labeling using several nucleolar protein markers. The fate of nucleolar XCAP-E was also followed when changes in the nucleoli morphology were artificially induced. The apparent exclusion of XCAP-E from the ribosomal DNA and its tight association with the granular component in all preparations suggest that it might be sequestrated in nucleoli during early stages of meiosis. Interestingly, both XCAP-D2 and XCAP-E were also detected in Cajal bodies, which are organelles suspected to play a role in the assembly/modification of the RNA transcription and processing machinery. The presence of two condensins in CBs might extend such a role of assembly to chromatin macromolecular components as well.
使用了几种抗体来检测两种凝聚素成员XCAP-E和XCAP-D2在非洲爪蟾卵母细胞核中的分布。发现XCAP-D2与灯刷染色体相关。含有XCAP-D2的染色体区域与高度浓缩染色质的结构域精确对应,这表明XCAP-D2在减数分裂染色质组织中起直接作用。相比之下,发现XCAP-E不在染色体上,但在核仁的颗粒成分中检测到高浓度。使用几种核仁蛋白标记物进行双重标记进一步证实了XCAP-E在核仁亚区的定位。当人为诱导核仁形态发生变化时,也追踪了核仁XCAP-E的命运。在所有制剂中,XCAP-E明显被排除在核糖体DNA之外,并与颗粒成分紧密结合,这表明它可能在减数分裂早期被隔离在核仁中。有趣的是,在卡哈尔体中也检测到了XCAP-D2和XCAP-E,卡哈尔体是一种怀疑在RNA转录和加工机器的组装/修饰中起作用的细胞器。两种凝聚素在卡哈尔体中的存在可能也将这种组装作用扩展到染色质大分子成分。