The positioning and dynamics of origins of replication in the budding yeast nucleus.
The positioning and dynamics of origins of replication in the budding yeast nucleus.
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DOI：
10.1083/jcb.152.2.385
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发表时间：
2001-01-22
期刊：
影响因子：
--
通讯作者：
Gasser SM
中科院分区：
文献类型：
--
作者：
Heun P;Laroche T;Raghuraman MK;Gasser SM
我们用荧光原位杂交和绿色荧光蛋白(GFP)标记的染色体结构域分析了完整酵母细胞中DNA复制的早期和晚期亚核位置。在这两种情况下，起始位置都是根据核孔染色或NUP49-GFP融合蛋白鉴定的核膜确定的。我们发现，在G1期，非端粒晚激发起源集中在紧邻核膜的区域，尽管这种定位在S相并不一定持续。相反，早期的放电起源在整个细胞周期中随机地定位在细胞核内。如果晚激发的端粒-近端起源在G1期从其染色体上下文中被切除，它仍然是晚激发的，但迅速远离它相关的端粒，这表明酵母染色体结构域的定位是高度动态的。对体内GFP标记来源的时间推移显微镜证实了这一点。我们认为，晚激发起点两侧的序列有助于将它们靶向G1相核的外围，在那里可以建立一种修改的染色质结构。修改后的染色质结构，反过来会延缓起源的放电，既是自主的，也是在细胞核内移动的。
We have analyzed the subnuclear position of early- and late-firing origins of DNA replication in intact yeast cells using fluorescence in situ hybridization and green fluorescent protein (GFP)–tagged chromosomal domains. In both cases, origin position was determined with respect to the nuclear envelope, as identified by nuclear pore staining or a NUP49-GFP fusion protein. We find that in G1 phase nontelomeric late-firing origins are enriched in a zone immediately adjacent to the nuclear envelope, although this localization does not necessarily persist in S phase. In contrast, early firing origins are randomly localized within the nucleus throughout the cell cycle. If a late-firing telomere-proximal origin is excised from its chromosomal context in G1 phase, it remains late-firing but moves rapidly away from the telomere with which it was associated, suggesting that the positioning of yeast chromosomal domains is highly dynamic. This is confirmed by time-lapse microscopy of GFP-tagged origins in vivo. We propose that sequences flanking late-firing origins help target them to the periphery of the G1-phase nucleus, where a modified chromatin structure can be established. The modified chromatin structure, which would in turn retard origin firing, is both autonomous and mobile within the nucleus.
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DOI：
10.1083/jcb.145.6.1119
发表时间：
1999-06-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Croft JA;Bridger JM;Boyle S;Perry P;Teague P;Bickmore WA
通讯作者：
Bickmore WA
影响因子：
2.1
作者：
Hayashi, A;Ogawa, H;Hiraoka, Y
通讯作者：
Hiraoka, Y
影响因子：
64.5
作者：
FERGUSON, BM;BREWER, BJ;FANGMAN, WL
通讯作者：
FANGMAN, WL
影响因子：
10.5
作者：
Friedman, KL;Diller, JD;Fangman, WL
通讯作者：
Fangman, WL
将蛋白质p270/TPR鉴定为核孔复合体核内细丝的组成部分。
DOI：
10.1083/jcb.136.3.515
发表时间：
1997-02-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Cordes VC;Reidenbach S;Rackwitz HR;Franke WW
通讯作者：
Franke WW