Human centromere repositioning “in progress”

Amor, David J.; Bentley, Karen; Ryan, Jacinta; Perry, Jo K.; Wong, Lee H.; Slater, Howard R.; Choo, K.H. Andy

doi:10.1073/pnas.0308637101

Cited by 200 publications

(221 citation statements)

References 46 publications

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“…We observed enrichment of RNAPII-ser2 at the active mitotic neocentromeres of both the mardel (10) and the PD-NC4 chromosomes ( Fig. 1 E and F) but not at the inactive alphoid centromere [marked by a faint CREST signal because of residual centromere protein B (CENP-B)] of PD-NC4 (16). These results suggest that RNAPII is localized to active kinetochores rather than satellite repeats and demonstrate an association between RNAPII localization at the kinetochore and centromere activity.…”

Section: Rnapii Localizes To the Mitotic Kinetochore And Is Associatementioning

confidence: 92%

“…All cells were cultured under standard conditions. Human 14ZBHT cells (16) were cultured with 100 μg/mL Zeocin (Invitrogen). Mouse ES monochromosomal cell hybrid ESmar10 cells (33) were cultured in DMEM with 15% FBS (vol/vol), 10 3 U/mL leukemic inhibitory factor, 0.1 mM β-mercaptoethanol, and 100 μg/mL Zeocin.…”

Section: Methodsmentioning

confidence: 99%

“…To establish whether RNAPII is associated with kinetochore function, we assayed for the presence of RNAPII-ser2 at the neocentromeres of two marker chromosomes: the human chromosome 10-derived mardel (10) marker chromosome containing a 10q25 neocentromere (15), and the pseudodicentric-neocentric chromosome 4 (PD-NC4), which contains an inactivated native α-satellite centromere and a functional neocentromere at 4q21 (16). We observed enrichment of RNAPII-ser2 at the active mitotic neocentromeres of both the mardel (10) and the PD-NC4 chromosomes ( Fig.…”

Section: Rnapii Localizes To the Mitotic Kinetochore And Is Associatementioning

confidence: 99%

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Active transcription and essential role of RNA polymerase II at the centromere during mitosis

Chan

Marshall

Saffery

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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241

287

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Transcription of the centromeric regions has been reported to occur in G1 and S phase in different species. Here, we investigate whether transcription also occurs and plays a functional role at the mammalian centromere during mitosis. We show the presence of actively transcribing RNA polymerase II (RNAPII) and its associated transcription factors, coupled with the production of centromere satellite transcripts at the mitotic kinetochore. Specific inhibition of RNAPII activity during mitosis leads to a decrease in centromeric α-satellite transcription and a concomitant increase in anaphase-lagging cells, with the lagging chromosomes showing reduced centromere protein C binding. These findings demonstrate an essential role of RNA-PII in the transcription of α-satellite DNA, binding of centromere protein C, and the proper functioning of the mitotic kinetochore.chromatin | noncoding RNA | epigenetics T he centromere is an essential chromosomal structure that mediates microtubule attachment during cell division to ensure correct chromosome segregation. Although centromere function is highly conserved, centromere DNA sequences show no evolutionary conservation. Instead, centromeric chromatin typically is filled with species-specific satellite DNA sequences that lack transcribed genes. The presence of functional ectopic centromeres (neocentromeres) at genomic regions devoid of classical satellite DNA repeats confirmed the epigenetic nature of centromere function (1).The centromere is organized into two broad domains characterized by distinct sets of epigenetic determinants. The centromere core domain comprises the centromere-specific histone H3 variant centromere protein A (CENP-A) that is essential for kinetochore formation, whereas the pericentric heterochromatin is vital for sister chromatid cohesion (for review, see ref.2). In yeast, pericentric outermost DNA repeats are transcribed and processed by RNAi machinery into siRNAs, which direct the deposition of heterochromatic markers such as H3K9me3 and HP1 at the pericentric heterochromatin (3). The RNAi pathway also has been shown to be vital for the establishment of pericentric heterochromatin in plant and animal cells (4, 5). However, although the depletion of Dicer in human-chicken hybrid cells causes defective pericentric heterochromatin, it does not affect the binding of CENP-A and centromere protein C (CENP-C) at the centromere core domain (6), suggesting that the RNAi pathway is not required for core centromere function. Our previous studies showed that transcription is permissible within the kinetochore domain of a human neocentromere (7, 8), but others have reported the presence of active genes within the rice kinetochore domain (9). Consistent with these reports, the CENP-A domain in Drosophila and human cells is enriched with the euchromatin-like marker H3K4me2 (10). Such studies suggest that transcription of centromeric chromatin is permissible and compatible with centromere function. Furthermore, we and others have demonstrated the presence of RNA at th...

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Section: Rnapii Localizes To the Mitotic Kinetochore And Is Associatementioning

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Rnapii Localizes To the Mitotic Kinetochore And Is Associatementioning

confidence: 99%

See 1 more Smart Citation

Active transcription and essential role of RNA polymerase II at the centromere during mitosis

Chan

Marshall

Saffery

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

241

287

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“…Eukaryotic centromeres, in general, comprise long DNA stretches, display little or no sequence consensus, and are epigenetically specified (20). Neo-centromeres can arise at chro-mosomal locales where none existed previously (37,38). Machineries for RNA interference and heterochromatin formation are important in the establishment of these "regional" centromeres (39).…”

Section: Discussionmentioning

confidence: 99%

Histone H3-variant Cse4-induced positive DNA supercoiling in the yeast plasmid has implications for a plasmid origin of a chromosome centromere

Huang

Chang

Cui

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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The Saccharomyces cerevisiae 2-μm plasmid is a multicopy selfish genome that resides in the nucleus. The genetic organization of the plasmid is optimized for stable, high-copy propagation in hostcell populations. The plasmid's partitioning system poaches host factors, including the centromere-specific histone H3-variant Cse4 and the cohesin complex, enabling replicated plasmid copies to segregate equally in a chromosome-coupled fashion. We have characterized the in vivo chromatin topology of the plasmid partitioning locus STB in its Cse4-associated and Cse4-nonassociated states. We find that the occupancy of Cse4 at STB induces positive DNA supercoiling, with a linking difference (ΔLk) contribution estimated between +1 and +2 units. One plausible explanation for this contrary topology is the presence of a specialized Cse4-containing nucleosome with a right-handed DNA writhe at a functional STB, contrasted by a standard histone H3-containing nucleosome with a left-handed DNA writhe at a nonfunctional STB. The similarities between STB and centromere in their nucleosome signature and DNA topology would be consistent with the potential origin of the unusual point centromere of budding yeast chromosomes from the partitioning locus of an ancestral plasmid.CEN evolution | nucleosome topology | reversome T he 2-μm plasmid of Saccharomyces cerevisiae resides in the nucleus at 40 to 60 copies per cell, and propagates itself with nearly the same stability as chromosomes (1, 2). The plasmid is a benign selfish DNA element that seems to provide no advantage to its host, but poses, at its normal copy number, no serious disadvantage either. In haploid cells the plasmid is organized into a cluster of three to five foci, and segregates as a cluster (3). This effective reduction in copy number necessitates an active partitioning system, comprised of two plasmid-coded proteins, Rep1 and Rep2, and a cis-acting partitioning locus STB, to ensure equal or nearly equal plasmid segregation. A decline in plasmid copy number because of rare missegregation events is corrected via DNA amplification triggered by the Flp sitespecific recombination system harbored by the plasmid (4, 5). Positive and negative regulatory circuits implemented through the plasmid-coded Raf1 protein and the Rep proteins ensure quick amplification response without the danger of runaway increase in copy number (6-8).The Rep-STB system channels several host factors involved in chromosome segregation toward the execution of plasmid segregation. These factors include the mitotic spindle, the spindleassociated motor Kip1, the RSC2 chromatin-remodeling complex, the centromere-specific histone H3-variant Cse4 (CenH3), and the yeast cohesin complex (9-15). The de novo assembly of the plasmid-partitioning complex at STB during the G1-S window of each cell cycle (9,12,14,16) is reminiscent of the assembly of the kinetochore complex at centromeres. However, kinetochore components have not been detected at STB by ChIP (13).The assembly of the plasmid-partitioning complex culmin...

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“…In one case, however, the chromosome was unstable (Rivera et al 1996), and in the other two cases, the neocentromere was embedded in the heterochromatic block of Yq. Very recently, Amor et al (2004) have reported a two-generation family, in which the active centromere of one chromosome 4 has been relocated to 4q21.3 in otherwise cytogenetically normal and mitotically stable karyotypes. Our case 2, and the case reported by Amor et al (2004) may be regarded as a present-day episode equivalent to new centromere appearance that occurred during the course of evolution.…”

Section: Human Neocentromeresmentioning

confidence: 99%

Recurrent Sites for New Centromere Seeding

et al. 2004

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Using comparative FISH and genomics, we have studied and compared the evolution of chromosome 3 in primates and two human neocentromere cases on the long arm of this chromosome. Our results show that one of the human neocentromere cases maps to the same 3q26 chromosomal region where a new centromere emerged in a common ancestor of the Old World monkeys ∼25-40 million years ago. Similarly, the locus in which a new centromere was seeded in the great apes' ancestor was orthologous to the site in which a new centromere emerged in the New World monkeys' ancestor. These data suggest the recurrent use of longstanding latent centromeres and that there is an inherent potential of these regions to form centromeres. The second human neocentromere case (3q24) revealed unprecedented features. The neocentromere emergence was not accompanied by any chromosomal rearrangement that usually triggers these events. Instead, it involved the functional inactivation of the normal centromere, and was present in an otherwise phenotypically normal individual who transmitted this unusual chromosome to the next generation. We propose that the formation of neocentromeres in humans and the emergence of new centromeres during the course of evolution share a common mechanism

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Human centromere repositioning “in progress”

Cited by 200 publications

References 46 publications

Active transcription and essential role of RNA polymerase II at the centromere during mitosis

Active transcription and essential role of RNA polymerase II at the centromere during mitosis

Histone H3-variant Cse4-induced positive DNA supercoiling in the yeast plasmid has implications for a plasmid origin of a chromosome centromere

Recurrent Sites for New Centromere Seeding

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