Cargando…
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...
Autores principales: | , |
---|---|
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 |
_version_ | 1783302635025072128 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5828391 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT thakurjitendra unexpectedconformationalvariationsofthehumancentromericchromatincomplex AT henikoffsteven unexpectedconformationalvariationsofthehumancentromericchromatincomplex |