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Unexpected conformational variations of the human centromeric chromatin complex

We combined classical salt fractionation with chromatin immunoprecipitation to recover human centromeric chromatin under native conditions. We found that >85% of the total centromeric chromatin is insoluble under conditions typically used for native chromatin extraction. To map both soluble and i...

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Detalles Bibliográficos
Autores principales: Thakur, Jitendra, Henikoff, Steven
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828391/
https://www.ncbi.nlm.nih.gov/pubmed/29386331
http://dx.doi.org/10.1101/gad.307736.117
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author Thakur, Jitendra
Henikoff, Steven
author_facet Thakur, Jitendra
Henikoff, Steven
author_sort Thakur, Jitendra
collection PubMed
description We combined classical salt fractionation with chromatin immunoprecipitation to recover human centromeric chromatin under native conditions. We found that >85% of the total centromeric chromatin is insoluble under conditions typically used for native chromatin extraction. To map both soluble and insoluble chromatin in situ, we combined CUT&RUN (cleavage under targets and release using nuclease), a targeted nuclease method, with salt fractionation. Using this approach, we observed unexpected structural and conformational variations of centromere protein A (CENP-A)-containing complexes on different α-satellite dimeric units within highly homogenous arrays. Our results suggest that slight α-satellite sequence differences control the structure and occupancy of the associated centromeric chromatin complex.
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spelling pubmed-58283912018-02-28 Unexpected conformational variations of the human centromeric chromatin complex Thakur, Jitendra Henikoff, Steven Genes Dev Research Communication We combined classical salt fractionation with chromatin immunoprecipitation to recover human centromeric chromatin under native conditions. We found that >85% of the total centromeric chromatin is insoluble under conditions typically used for native chromatin extraction. To map both soluble and insoluble chromatin in situ, we combined CUT&RUN (cleavage under targets and release using nuclease), a targeted nuclease method, with salt fractionation. Using this approach, we observed unexpected structural and conformational variations of centromere protein A (CENP-A)-containing complexes on different α-satellite dimeric units within highly homogenous arrays. Our results suggest that slight α-satellite sequence differences control the structure and occupancy of the associated centromeric chromatin complex. Cold Spring Harbor Laboratory Press 2018-01-01 /pmc/articles/PMC5828391/ /pubmed/29386331 http://dx.doi.org/10.1101/gad.307736.117 Text en © 2018 Thakur and Henikoff; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by/4.0/ This article, published in Genes & Development, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.
spellingShingle Research Communication
Thakur, Jitendra
Henikoff, Steven
Unexpected conformational variations of the human centromeric chromatin complex
title Unexpected conformational variations of the human centromeric chromatin complex
title_full Unexpected conformational variations of the human centromeric chromatin complex
title_fullStr Unexpected conformational variations of the human centromeric chromatin complex
title_full_unstemmed Unexpected conformational variations of the human centromeric chromatin complex
title_short Unexpected conformational variations of the human centromeric chromatin complex
title_sort unexpected conformational variations of the human centromeric chromatin complex
topic Research Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828391/
https://www.ncbi.nlm.nih.gov/pubmed/29386331
http://dx.doi.org/10.1101/gad.307736.117
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