H3K9me3 maintenance on a human artificial chromosome is required for segregation but not centromere epigenetic memory

Martins, Nuno M. C.; Cisneros-Soberanis, Fernanda; Pesenti, Elisa; Kochanova, Natalia Y.; Shang, Wei-Hao; Hori, Tomohide; Nagase, Takahiro; Kimurâ, Hiroshi; Larionov, Vladimir; Masumoto, Hiroshi; Fukagawa, Tatsuo; Earnshaw, William C.

doi:10.1242/jcs.242610

Cited by 15 publications

(21 citation statements)

References 129 publications

(181 reference statements)

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“…Nevertheless, neocentromeres and ectopic CENP-A sites are not randomly distributed across the human genome [ 149 ], implying that certain loci have centromeric potential, whereas other loci do not. Studies on human artificial chromosomes showed that a chromatin environment containing a distinct set and distribution of histone modifications was important for CENP-A chromatin formation as well [ 162 , 163 , 164 ]. Using the marker chromosome mardel(10), L1 retrotransposon transcripts have been found to facilitate the formation of CENP-A chromatin [ 165 ].…”

Section: Centromeric Transcription In Diseasementioning

confidence: 99%

Centromeric Transcription: A Conserved Swiss-Army Knife

Arunkumar

Melters

2020

Genes

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In most species, the centromere is comprised of repetitive DNA sequences, which rapidly evolve. Paradoxically, centromeres fulfill an essential function during mitosis, as they are the chromosomal sites wherein, through the kinetochore, the mitotic spindles bind. It is now generally accepted that centromeres are transcribed, and that such transcription is associated with a broad range of functions. More than a decade of work on this topic has shown that centromeric transcripts are found across the eukaryotic tree and associate with heterochromatin formation, chromatin structure, kinetochore structure, centromeric protein loading, and inner centromere signaling. In this review, we discuss the conservation of small and long non-coding centromeric RNAs, their associations with various centromeric functions, and their potential roles in disease.

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Section: Centromeric Transcription In Diseasementioning

confidence: 99%

Centromeric Transcription: A Conserved Swiss-Army Knife

Arunkumar

Melters

2020

Genes

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“…This binding therefore facilitates complete resolution of catenated loci, particularly CEN loci (for preventing UFBs) and chromosome bridges. Martins et al, 2020) and consistent with this, it is known that UFBs form when catenated CEN DNA molecules fail to be resolved before anaphase (Ke et al, 2011;Nielsen et al, 2015). Marking of catenated CEN loci with cues such as H3K27me3 is therefore a key mechanism that recruits TopoIIa to ensure resolution of the catenated CEN loci in a timely manner, before or during anaphase.…”

Section: Discussionmentioning

confidence: 58%

Role of the Topoisomerase IIα Chromatin Tether domain in Nucleosome Binding & Chromosome Segregation

Sundararajan

Park

Johansson

et al. 2021

Preprint

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Due to the intrinsic nature of DNA replication, replicated genomes retain catenated genomic loci that must be resolved to ensure faithful segregation of sister chromatids in mitosis. Type II DNA Topoisomerase (TopoII) decatenates the catenated genomic DNA through its unique Strand Passage Reaction (SPR). Loss of SPR activity results in anaphase chromosome bridges and formation of Polo-like Kinase Interacting Checkpoint Helicase (PICH)-coated ultra-fine DNA bridges (UFBs) whose timely resolution is required to prevent micronuclei formation. Vertebrates have two TopoII isoforms– TopoIIα and TopoIIβ, that share a conserved catalytic core. However, the essential mitotic function of TopoIIα cannot be compensated by TopoIIβ, due to differences in their catalytically inert C-terminal domains (CTDs). Using genome-edited human cells, we show that specific binding of TopoIIα to methylated histone, tri-methylated H3K27 (H3K27me3), via its Chromatin Tether (ChT) domain within the CTD contributes critically to avoid anaphase UFB formation. Reducing H3K27 methylation prior to mitosis increases UFBs, revealing a requirement for proper establishment of H3K27me3 after DNA replication to facilitate TopoIIα-ChT dependent UFB prevention. We propose that interaction of the TopoIIα-ChT with H3K27me3 is a key factor that ensures the complete resolution of catenated loci to permit faithful chromosome segregation in human cells.

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“…This is probably because the percentage of mis-segregating cells is determined by scoring individual cells in which the level of CENP-A falls below a critical threshold, and it is not determined by the average CENP-A level in the cell population, as already described. 40 …”

Section: Resultsmentioning

confidence: 99%

Analysis of Complex DNA Rearrangements during Early Stages of HAC Formation

et al. 2020

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Human artificial chromosomes (HACs) are important tools for epigenetic engineering, for measuring chromosome instability (CIN), and for possible gene therapy. However, their use in the latter is potentially limited because the input HAC-seeding DNA can undergo an unpredictable series of rearrangements during HAC formation. As a result, after transfection and HAC formation, each cell clone contains a HAC with a unique structure that cannot be precisely predicted from the structure of the HAC-seeding DNA. Although it has been reported that these rearrangements can happen, the timing and mechanism of their formation has yet to be described. Here we synthesized a HAC-seeding DNA with two distinct structural domains and introduced it into HT1080 cells. We characterized a number of HAC-containing clones and subclones to track DNA rearrangements during HAC establishment. We demonstrated that rearrangements can occur early during HAC formation. Subsequently, the established HAC genomic organization is stably maintained across many cell generations. Thus, early stages in HAC formation appear to at least occasionally involve a process of DNA shredding and shuffling that resembles chromothripsis, an important hallmark of many cancer types. Understanding these events during HAC formation has critical implications for future efforts aimed at synthesizing and exploiting synthetic human chromosomes.

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H3K9me3 maintenance on a human artificial chromosome is required for segregation but not centromere epigenetic memory

Cited by 15 publications

References 129 publications

Centromeric Transcription: A Conserved Swiss-Army Knife

Centromeric Transcription: A Conserved Swiss-Army Knife

Role of the Topoisomerase IIα Chromatin Tether domain in Nucleosome Binding & Chromosome Segregation

Analysis of Complex DNA Rearrangements during Early Stages of HAC Formation

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