Dataset Information

Structural Basis for CAL1-Mediated Centromere Maintenance

ABSTRACT: Intermolecular interactions stabilising the CAL1–CENP-A/H4 complex were analysed using chemical Cross-Linking Mass Spectrometry (CLMS). Purified recombinant CAL11-160–CENP-A101-225–H4 complex was crosslinked using EDC and BS3. The crosslinked peptides were analysed by mass spectrometry to identify intra- and intermolecular contacts. Notably, the data revealed several intramolecular crosslinks between the N- and C-terminal regions of CAL11-160, suggesting a direct interaction between these regions.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

SUBMITTER: Juan Zou

LAB HEAD: Juri Rappsilber

PROVIDER: PXD017238 | Pride | 2020-03-11

REPOSITORIES: Pride

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Publications

Structural basis for centromere maintenance by Drosophila CENP-A chaperone CAL1.

Medina-Pritchard Bethan B Lazou Vasiliki V Zou Juan J Byron Olwyn O Abad Maria A MA Rappsilber Juri J Heun Patrick P Jeyaprakash A Arockia AA

The EMBO journal 20200305 7

Centromeres are microtubule attachment sites on chromosomes defined by the enrichment of histone variant CENP-A-containing nucleosomes. To preserve centromere identity, CENP-A must be escorted to centromeres by a CENP-A-specific chaperone for deposition. Despite this essential requirement, many eukaryotes differ in the composition of players involved in centromere maintenance, highlighting the plasticity of this process. In humans, CENP-A recognition and centromere targeting are achieved by HJUR ...[more]

PMID: 32134144

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Project description:Specialized chromatin containing CENP-A nucleosomes instead of H3 nucleosomes is found at all centromeres. However, the mechanisms that specify the locations at which CENP-A chromatin is assembled remain elusive in organisms with regional, epigenetically regulated centromeres. It is known that normal centromeric DNA is transcribed in several systems including the fission yeast, Schizosaccharomyces pombe. Here, we show that factors which preserve stable histone H3 chromatin during transcription also play a role in preventing promiscuous CENP-A(Cnp1) deposition in fission yeast. Mutations in the histone chaperone FACT impair the maintenance of H3 chromatin on transcribed regions and promote widespread CENP-A(Cnp1) incorporation at non-centromeric sites. FACT has little or no effect on CENP-A(Cnp1) assembly at endogenous centromeres where CENP-A(Cnp1) is normally assembled. In contrast, Clr6 complex II (Clr6-CII; equivalent to Rpd3S) histone deacetylase function has a more subtle impact on the stability of transcribed H3 chromatin and acts to prevent the ectopic accumulation of CENP-A(Cnp1) at specific loci, including subtelomeric regions, where CENP-A(Cnp1) is preferentially assembled. Moreover, defective Clr6-CII function allows the de novo assembly of CENP-A(Cnp1) chromatin on centromeric DNA, bypassing the normal requirement for heterochromatin. Thus, our analyses show that alterations in the process of chromatin assembly during transcription can destabilize H3 nucleosomes and thereby allow CENP-A(Cnp1) to assemble in its place. We propose that normal centromeres provide a specific chromatin context that limits reassembly of H3 chromatin during transcription and thereby promotes the establishment of CENP-A(Cnp1) chromatin and associated kinetochores. These findings have important implications for genetic and epigenetic processes involved in centromere specification. In total, 24 samples: 22 ChIP DNA files (10 different conditions), 2 Input files.

Dataset Information

Structural Basis for CAL1-Mediated Centromere Maintenance

Publications

Structural basis for centromere maintenance by Drosophila CENP-A chaperone CAL1.

Similar Datasets

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