Developmental regulation of Drosophila DNA topoisomerase II.
Developmental regulation of Drosophila DNA topoisomerase II.
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果蝇 DNA 拓扑异构酶 II 的发育调控。
DOI：
10.1083/jcb.112.2.203
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Fisher,PA
中科院分区：
文献类型：
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作者：
Whalen,AM;McConnell,M;Fisher,PA
亲和纯化的多克隆抗体被用来定量DNA拓扑异构酶II(TOPO II)在整个果蝇发育过程中的稳态水平。尽管在不同的阶段记录到了广泛的波动，但这些波动与核层粘连蛋白水平的变化是平行的，核层粘连蛋白是一种用作内部标准的核结构蛋白。这种情况的例外是成年男性，他们的Lamin水平相对于topo II显著升高。Topo II和lamin mRNA的Northern印迹分析与蛋白质的免疫印迹分析相结合，揭示了这两种不同信息的水平波动，这两种不同的信息与彼此及其各自的翻译产物的变化平行。生化和免疫化学分析由间接免疫荧光和原位免疫过氧化物酶实验补充。Topo II在所研究的大多数但不是所有类型的细胞中分布于细胞核。果蝇TOPO II的这些结果显然与其他脊椎动物系统工作人员的结果不一致(例如，参见Heck、M.M.S.和W.C.Earnshaw)。1986年。J.细胞生物学。103：2569-2581；Heck，M.M.S.，W.N.Hittelman，W.C.Earnshaw。1988年。程序娜塔莉。阿卡德。SCI。美国。85：1086-1090)，提示在果蝇中，Topo II可能不是一个有用的增殖状态的标志。
Affinity-purified polyclonal antibodies were used to quantitate steady-state levels of DNA topoisomerase II (topo II) throughout Drosophila development. Although wide fluctuations were recorded at different stages, these fluctuations were paralleled by changes in levels of the nuclear lamin, a nuclear structural protein used as an internal standard. The exception to this was adult males where lamin levels were significantly elevated relative to topo II. Northern blot analyses of topo II and lamin mRNA, performed in conjunction with immunoblot analyses of protein revealed fluctuations in levels of the two different messages that paralleled changes in each other and in their respective translation products. Biochemical and immunochemical analyses were complemented by indirect immunofluorescence and immunoperoxidase experiments performed in situ. topo II was found distributed throughout nuclei in most but not all cell types examined. These results for Drosophila topo II are apparently at odds with those obtained by others working in vertebrate systems (see, for example, Heck, M. M. S. and W. C. Earnshaw. 1986. J. Cell Biol. 103:2569-2581; Heck, M. M. S., W. N. Hittelman, and W. C. Earnshaw. 1988. Proc. Natl. Acad. Sci. USA. 85:1086-1090) and suggest that in Drosophila, topo II may not be a useful marker for the proliferative state.