Transcription within a Functional Human Centromere

Saffery, Richard; Sümer, Hüseyin; Hassan, Sara; Wong, Lee H.; Craig, Jeffrey M; Todokoro, Kazuo; Anderson, Melissa A.; Stafford, Angela J.; Choo, K.H. Andy

doi:10.1016/s1097-2765(03)00279-x

Cited by 137 publications

(168 citation statements)

References 35 publications

(4 reference statements)

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“…This is consistent with studies of the role of transcription and RNAi in centromere activity in S. pombe (Volpe et al, 2002(Volpe et al, , 2003Chen et al, 2008); however, the role for transcription in metazoan kinetochore activity has been little studied (Saffery et al, 2003;Fukagawa et al, 2004;Wong et al, 2007;Chueh et al, 2009). Our previous studies found that alphoid tetO kinetochore inactivation by the tTS correlated with the recruitment of heterochromatin protein HP1␣ and that direct recruitment of HP1␣ as a TetR:EYFP fusion protein could also inactivate the kinetochore (Nakano et al, 2008).…”

Section: Molecular Biology Of the Cell 4200supporting

confidence: 90%

Hierarchical Inactivation of a Synthetic Human Kinetochore by a Chromatin Modifier

Cardinale

Bergmann

Kelly

et al. 2009

MBoC

114

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We previously used a human artificial chromosome (HAC) with a synthetic kinetochore that could be targeted with chromatin modifiers fused to tetracycline repressor to show that targeting of the transcriptional repressor tTS within kinetochore chromatin disrupts kinetochore structure and function. Here we show that the transcriptional corepressor KAP1, a downstream effector of the tTS, can also inactivate the kinetochore. The disruption of kinetochore structure by KAP1 subdomains does not simply result from loss of centromeric CENP-A nucleosomes. Instead it reflects a hierarchical disruption of the outer kinetochore, with CENP-C levels falling before CENP-A levels and, in certain instances, CENP-H being lost more readily than CENP-C. These results suggest that this novel approach to kinetochore dissection may reveal new patterns of protein interactions within the kinetochore. INTRODUCTIONThe centromere/kinetochore is one of the most complex cellular substructures, with more than 80 protein components described to date (reviewed in Carroll and Straight, 2006;Cheeseman and Desai, 2008;Fukagawa, 2008;Vagnarelli et al., 2008). These components perform the complex job of attaching chromosomes to the mitotic spindle; ensuring that those attachments are correct; signaling to delay mitotic progression if they are not, and regulating the movements of the chromosomes toward the spindle poles in anaphase.The kinetochore is assembled at a unique locus on each natural chromosome. However, for organisms with regional centromeres (Pluta et al., 1995), this reflects only a preference¤, and not an absolute requirement for particular DNA sequences. Kinetochores can form on a wide range of singlecopy and repeated DNA sequences, leading to the conclusion that the ultimate determinants of kinetochore assembly are epigenetic. The long-term purpose of our studies is to determine the epigenetic "landscape" that promotes kinetochore assembly and its maintenance during cell divisions.Experiments including yeast genetics, RNA interference (RNAi) studies in mammalian cells, and gene knockout analysis in mouse and chicken DT40 cells have revealed that kinetochores assemble on a foundation of specialized chromatin containing the kinetochore-specific histone H3 variant CENP-A (Earnshaw and Rothfield, 1985). CENP-A is upstream of almost all other known components in the kinetochore assembly pathway. However, that pathway is multiplex, as recent studies in chicken and Drosophila show that inner kinetochore proteins CENP-H and -C are required for normal CENP-A loading or retention (Okada et al., 2006;Goshima et al., 2007;Erhardt et al., 2008).Our work was inspired by an approach first developed a number of years ago in which cloned fragments of human centromeric DNA were used to form human artificial chromosomes (HACs) in HT1080 fibrosarcoma cells (Harrington et al., 1997;Ikeno et al., 1998). Originally, HAC formation was only achieved with regular repeated arrays of ␣-satellite DNA containing CENP-B boxes Ohzeki et al., 2002;Okada et al., 2007)....

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Section: Molecular Biology Of the Cell 4200supporting

confidence: 90%

Hierarchical Inactivation of a Synthetic Human Kinetochore by a Chromatin Modifier

Cardinale

Bergmann

Kelly

et al. 2009

MBoC

114

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“…A more recent study has shown that centrochromatin has a histone modification pattern resembling that found in the downstream body of transcribed genes (Bergmann et al, 2011). Indeed, previous studies have detected transcription within rice centromeres (Nagaki et al, 2004), human neocentromeres (Saffery et al, 2003) and also within the alphoid array of a derivative of the synthetic HAC used in the present study (Iida et al, 2010). The chromatin profile at active kinetochores resembles the 'yellow' chromatin described in a recent paper (Filion et al, 2010).…”

Section: Discussionsupporting

confidence: 77%

Epigenetic engineering: histone H3K9 acetylation is compatible with kinetochore structure and function

Bergmann

Jakubsche

Martins

et al. 2012

Journal of Cell Science

103

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SummaryHuman kinetochores are transcriptionally active, producing very low levels of transcripts of the underlying alpha-satellite DNA. However, it is not known whether kinetochores can tolerate acetylated chromatin and the levels of transcription that are characteristic of housekeeping genes, or whether kinetochore-associated 'centrochromatin', despite being transcribed at a low level, is essentially a form of repressive chromatin. Here, we have engineered two types of acetylated chromatin within the centromere of a synthetic human artificial chromosome. Tethering a minimal NF-kB p65 activation domain within kinetochore-associated chromatin produced chromatin with high levels of histone H3 acetylated on lysine 9 (H3K9ac) and an ,10-fold elevation in transcript levels, but had no substantial effect on kinetochore assembly or function. By contrast, tethering the herpes virus VP16 activation domain produced similar modifications in the chromatin but resulted in an ,150-fold elevation in transcripts, approaching the level of transcription of an endogenous housekeeping gene. This rapidly inactivated kinetochores, causing a loss of assembled CENP-A and blocking further CENP-A assembly. Our data reveal that functional centromeres in vivo show a remarkable plasticity -kinetochores tolerate profound changes to their chromatin environment, but appear to be critically sensitive to the level of centromeric transcription.

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“…While the pericentromere itself is important for providing a boundary around the centromere core, another critical region in maintaining chromosome integrity is the chromatin barrier between the pericentromere and the centromere (Saffery et al 2003;Scott et al 2007). In fission yeast, nucleosome-free tRNA act as a barrier preventing the spreading of heterochromatin into the centromere (Scott et al 2007) (Fig.…”

Section: Boundary Elementsmentioning

confidence: 99%

Pericentric and centromeric transcription: a perfect balance required

2012

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The pericentromere and centromere regions of the genome have previously been considered tightly compacted and transcriptionally inert. However, there is mounting evidence that these regions not only actively produce transcripts but that these pericentric and centromeric transcripts are also vital to maintaining genome stability and proper cell division. In this review, we define the pericentromere and centromere of eukaryotic chromosomes in terms of their histone modifications and their nascent transcripts. In addition, we present the currently known roles these transcripts play in heterochromatin formation, development, and differentiation, as well as their interaction with centromeric proteins, and ultimately centromere function. Recent work has added considerable complexity to the theoretical framework defining the innate requirement for pericentric and centromeric transcription. It is clear that maintaining a fine balance of transcriptional output is critical, as deviations from this balance result in centromere disfunction and genomic instability.

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Transcription within a Functional Human Centromere

Cited by 137 publications

References 35 publications

Hierarchical Inactivation of a Synthetic Human Kinetochore by a Chromatin Modifier

Hierarchical Inactivation of a Synthetic Human Kinetochore by a Chromatin Modifier

Epigenetic engineering: histone H3K9 acetylation is compatible with kinetochore structure and function

Pericentric and centromeric transcription: a perfect balance required

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