Mitotic segregation of viral and cellular acentric extrachromosomal molecules by chromosome tethering.
Mitotic segregation of viral and cellular acentric extrachromosomal molecules by chromosome tethering.
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通过染色体束缚进行病毒和细胞无着丝粒染色体外分子的有丝分裂分离。
DOI：
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发表时间：
2001
影响因子：
4
通讯作者：
Geoffrey M. Wahl
中科院分区：
文献类型：
--
作者：
Teru Kanda;Michèle Otter;Geoffrey M. Wahl
有丝分裂染色体分离是由附着在着丝粒上的纺锤体微管介导的。然而，最近的研究表明，无着丝粒DNA分子，如病毒复制子和双微小染色体，可以有效地分离成子细胞与有丝分裂染色体。基于病毒和细胞无着丝粒分子之间的这种相似性，我们将EB病毒载体引入具有双微小染色体的细胞中，并比较它们的有丝分裂行为。我们将lac操纵基因重复序列添加到EB病毒载体中，这使我们能够容易地在表达lac阻遏物和绿色荧光蛋白之间的融合蛋白的细胞中鉴定转基因。出乎意料的是，我们发现EB病毒载体整合到无着丝粒双微小染色体，但不进入正常染色体，在所有六个稳定转染的克隆检查。瞬时转染的EB病毒载体与轮状前中期染色体嵌合体随机结合，而稳定转染克隆中的双微小染色体与EB病毒载体的嵌合体则始终附着在染色体嵌合体的周围。这些嵌合无着丝粒分子忠实地代表了天然双微染色体的行为，为分析它们在整个细胞周期中在活细胞中的行为提供了工具。进一步的详细分析，包括实时观察，揭示了双微小染色体似乎被排斥在纺锤体两极的同时，它们附着在染色体外周，而着丝粒区域被附着的微管拉向两极。破坏微管组织消除了这种周边定位的双微小染色体，但它并不影响他们与染色体。结果表明，在一个模型中，双微小染色体，而不是EB病毒载体，受到微管介导的反极性力，而它们都采用染色体拴系策略，以增加其分离到子细胞。
Mitotic chromosome segregation is mediated by spindle microtubules attached to centromeres. Recent studies, however, revealed that acentric DNA molecules, such as viral replicons and double minute chromosomes, can efficiently segregate into daughter cells by associating with mitotic chromosomes. Based on this similarity between viral and cellular acentric molecules, we introduced Epstein-Barr virus vectors into cells harboring double minute chromosomes and compared their mitotic behaviors. We added lac operator repeats to an Epstein-Barr virus vector, which enabled us to readily identify the transgene in cells expressing a fusion protein between the lac repressor and green fluorescent protein. Unexpectedly, we found that Epstein-Barr virus vectors integrated into the acentric double minute chromosomes, but not into normal chromosomes, in all of the six stably transfected clones examined. While transiently transfected Epstein-Barr virus vectors randomly associated with wheel-shaped prometaphase chromosome rosettes, the chimeras of double minute chromosomes and Epstein-Barr virus vectors in stably transfected clones always attached to the periphery of chromosome rosettes. These chimeric acentric molecules faithfully represented the behavior of native double minute chromosomes, providing a tool for analyzing their behavior in living cells throughout the cell cycle. Further detailed analyses, including real-time observations, revealed that double minute chromosomes appeared to be repelled from the spindle poles at the same time that they attached to the chromosome periphery, while centromeric regions were pulled poleward by the attached microtubules. Disrupting microtubule organization eliminated such peripheral localization of double minute chromosomes, but it did not affect their association with chromosomes. The results suggest a model in which double minute chromosomes, but not Epstein-Barr virus vectors, are subject to the microtubule-mediated antipolar force, while they both employ chromosome tethering strategies to increase their segregation to daughter cells.
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DOI：
10.1016/0165-1110(92)90010-7
发表时间：
1992
期刊：
Mutation research
影响因子：
--
作者：
Kimmel,M;Axelrod,DE;Wahl,GM
通讯作者：
Wahl,GM
影响因子：
3.7
作者：
PETTI, L;SAMPLE, C;KIEFF, E
通讯作者：
KIEFF, E
影响因子：
11.2
作者：
Geoffrey M. Wahl
通讯作者：
Geoffrey M. Wahl
使用 lac 操纵子/lac 阻遏物报告系统对大规模染色质结构和动力学进行可视化。
影响因子：
--
作者：
Belmont,AS;Li,G;Sudlow,G;Robinett,C
通讯作者：
Robinett,C
与核基质的附着介导染色质结构的特定改变。
DOI：
10.1073/pnas.95.25.14757
发表时间：
1998
影响因子：
11.1
作者：
Pemov,A;Bavykin,S;Hamlin,JL
通讯作者：
Hamlin,JL