Chromosome engineering allows the efficient isolation of vertebrate neocentromeres.
Chromosome engineering allows the efficient isolation of vertebrate neocentromeres.
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DOI：
10.1016/j.devcel.2013.02.009
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发表时间：
2013-03-25
影响因子：
11.8
通讯作者：
Fukagawa, Tatsuo
中科院分区：
文献类型：
--
作者：
Shang, Wei-Hao;Hori, Tetsuya;Martins, Nuno M. C.;Toyoda, Atsushi;Misu, Sadahiko;Monma, Norikazu;Hiratani, Ichiro;Maeshima, Kazuhiro;Ikeo, Kazuho;Fujiyama, Asao;Kimura, Hiroshi;Earnshaw, William C.;Fukagawa, Tatsuo
在大多数生物中，着丝粒是由序列独立的表观遗传机制指定的。很少，着丝粒重新定位的结果在异位部位形成新着丝粒。然而，控制新着丝粒如何以及在何处形成的机制尚不清楚。在这里，我们在鸡DT 40细胞中建立了一个染色体工程系统，使我们能够有效地分离含有新着丝粒的染色体。新着丝粒似乎在结构和功能上等同于天然着丝粒。用18个新着丝粒进行的染色质免疫沉淀测序（ChIP-seq）分析显示，着丝粒特异性组蛋白H3变体CENP-A在每个新着丝粒处占据约40 kb的区域，其对特定的DNA序列基序没有偏好。此外，我们发现新着丝粒与组蛋白修饰H3 K9 me 3、H3 K4 me 2和H3 K36 me 3或早期复制时间无关。重要的是，在内源性着丝粒周围检测到低但显著水平的CENP-A，如果去除着丝粒核心，其能够接种新着丝粒组装。总之，我们的实验系统为了解新着丝粒的形成提供了有价值的见解。染色体工程在鸡DT 40细胞中有效地产生新着丝粒。CENP-A在每个新着丝粒处可重复地占据约40 kb的基因组区域。非动粒CENP-A似乎起着新着丝粒组装的种子的作用。Shang等人在DT 40细胞中产生了鸡新着丝粒的集合。他们的分析表明，新着丝粒的形成与预期的组蛋白修饰或复制时间无关，而是取决于组蛋白H3变体CENP-A的种子组装。
Centromeres are specified by sequence-independent epigenetic mechanisms in most organisms. Rarely, centromere repositioning results in neocentromere formation at ectopic sites. However, the mechanisms governing how and where neocentromeres form are unknown. Here, we established a chromosome-engineering system in chicken DT40 cells that allowed us to efficiently isolate neocentromere-containing chromosomes. Neocentromeres appear to be structurally and functionally equivalent to native centromeres. Chromatin immunoprecipitation sequencing (ChIP-seq) analysis with 18 neocentromeres revealed that the centromere-specific histone H3 variant CENP-A occupies an ∼40 kb region at each neocentromere, which has no preference for specific DNA sequence motifs. Furthermore, we found that neocentromeres were not associated with histone modifications H3K9me3, H3K4me2, and H3K36me3 or with early replication timing. Importantly, low but significant levels of CENP-A are detected around endogenous centromeres, which are capable of seeding neocentromere assembly if the centromere core is removed. In summary, our experimental system provides valuable insights for understanding how neocentromeres form. ► Chromosome engineering efficiently generates neocentromeres in chicken DT40 cells ► CENP-A reproducibly occupies an ∼40 kb genomic region at each neocentromere ► Nonkinetochore CENP-A appears to function as a seed for neocentromere assembly Centromeres are specified by sequence-independent epigenetic mechanisms. Shang et al. generated a collection of chicken neocentromeres in DT40 cells. Their analysis indicates that neocentromere formation does not correlate with the expected histone modifications or with replication timing, but rather depends on the histone H3 variant CENP-A to seed assembly.
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DOI：
10.1073/pnas.0507947103
发表时间：
2006-03-14
影响因子：
11.1
作者：
Lam, AL;Boivin, CD;Sullivan, BA
通讯作者：
Sullivan, BA
影响因子：
64.5
作者：
Hori, Tetsuya;Amano, Miho;Fukagawa, Tatsuo
通讯作者：
Fukagawa, Tatsuo
DOI：
10.1083/jcb.201001035
发表时间：
2010-07-26
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bassett EA;Wood S;Salimian KJ;Ajith S;Foltz DR;Black BE
通讯作者：
Black BE
影响因子：
3.3
作者：
Hori, Tetsuya;Okada, Masahiro;Fukagawa, Tatsuo
通讯作者：
Fukagawa, Tatsuo
影响因子：
2.6
作者：
Jin, Weiwei;Lamb, Jonathan C.;Jiang, Jiming
通讯作者：
Jiang, Jiming