期刊
GENES & DEVELOPMENT
卷 32, 期 1, 页码 20-25出版社
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1101/gad.307736.117
关键词
centromeres; chromatin; methodology
资金
- NIH HHS [S10 OD020069] Funding Source: Medline
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 alpha-satellite dimeric units within highly homogenous arrays. Our results suggest that slight alpha-satellite sequence differences control the structure and occupancy of the associated centromeric chromatin complex.
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