Iwabata Kazuki, Sakaguchi Kengo
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba-ken 278-8510, Japan.
Chromosoma. 2008 Jun;117(3):297-302. doi: 10.1007/s00412-008-0149-5. Epub 2008 Feb 15.
We reported previously that Coprinus cinereus Lim15/Dmc1 (CcLim15), a meiosis-specific recA-like protein, could specifically activate C. cinereus DNA topoisomerase II (CcTopII). In particular, it enhanced the catenation activity of CcTopII in vitro at the meiotic prophase stage (Iwabata K, Koshiyama A, Yamaguchi T, Sugawara H, Hamada NF, Namekawa HS, Ishii S, Ishizaki T, Chiku H, Nara T, Sakaguchi K, Nucleic Acids Res, 33:5809-5818, 2005). In this study, the interaction between CcTopII and CcLim15, especially during catenation, was investigated in detail using atomic force microscopy. We demonstrated earlier that CcLim15 enhanced the catenation activity of CcTopII in a dose-dependent manner. When using two different-sized plasmid rings (5.4 and 3 kbp), which did not have any homologous sequence regions, equal proportions of homologous and heterologous catenanes were produced, suggesting that CcLim15 causes an increase in catenation activity irrespective of the presence of homologous sequences between the rings. We also showed that CcLim15 works as a DNA-condensing agent. Therefore, we speculate that CcLim15 may work as a DNA-condensing factor specific to the zygotene event and that CcTopII is likely to resolve tangles when the chromosomes initiate pairing at multiple sites by CcLim15.
我们之前报道过,灰盖鬼伞的减数分裂特异性类recA蛋白Coprinus cinereus Lim15/Dmc1(CcLim15)能够特异性激活灰盖鬼伞DNA拓扑异构酶II(CcTopII)。特别是,它在减数分裂前期增强了CcTopII在体外的连环化活性(岩田K、小石山A、山口T、菅原H、滨田NF、名川HS、石井S、石崎T、知久H、奈良T、坂口K,《核酸研究》,33:5809 - 5818,2005年)。在本研究中,使用原子力显微镜详细研究了CcTopII与CcLim15之间的相互作用,尤其是在连环化过程中的相互作用。我们之前证明了CcLim15以剂量依赖的方式增强了CcTopII的连环化活性。当使用两个没有任何同源序列区域的不同大小的质粒环(5.4和3 kbp)时,产生了相等比例的同源和异源连环体,这表明无论环之间是否存在同源序列,CcLim15都会导致连环化活性增加。我们还表明CcLim15作为一种DNA凝聚剂发挥作用。因此,我们推测CcLim15可能作为一种特异性作用于偶线期事件的DNA凝聚因子发挥作用,并且当染色体通过CcLim15在多个位点开始配对时,CcTopII可能会解开缠结。