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Steven Henikoff

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https://www.readbyqxmd.com/read/29651053/targeted-in-situ-genome-wide-profiling-with-high-efficiency-for-low-cell-numbers
#1
Peter J Skene, Jorja G Henikoff, Steven Henikoff
Cleavage under targets and release using nuclease (CUT&RUN) is an epigenomic profiling strategy in which antibody-targeted controlled cleavage by micrococcal nuclease releases specific protein-DNA complexes into the supernatant for paired-end DNA sequencing. As only the targeted fragments enter into solution, and the vast majority of DNA is left behind, CUT&RUN has exceptionally low background levels. CUT&RUN outperforms the most widely used chromatin immunoprecipitation (ChIP) protocols in resolution, signal-to-noise ratio and depth of sequencing required...
May 2018: Nature Protocols
https://www.readbyqxmd.com/read/29426353/precise-genome-wide-mapping-of-single-nucleosomes-and-linkers-in-vivo
#2
Răzvan V Chereji, Srinivas Ramachandran, Terri D Bryson, Steven Henikoff
We developed a chemical cleavage method that releases single nucleosome dyad-containing fragments, allowing us to precisely map both single nucleosomes and linkers with high accuracy genome-wide in yeast. Our single nucleosome positioning data reveal that nucleosomes occupy preferred positions that differ by integral multiples of the DNA helical repeat. By comparing nucleosome dyad positioning maps to existing genomic and transcriptomic data, we evaluated the contributions of sequence, transcription, and histones H1 and H2A...
February 9, 2018: Genome Biology
https://www.readbyqxmd.com/read/29386331/unexpected-conformational-variations-of-the-human-centromeric-chromatin-complex
#3
Jitendra Thakur, Steven Henikoff
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...
January 1, 2018: Genes & Development
https://www.readbyqxmd.com/read/29365169/non-b-form-dna-is-enriched-at-centromeres
#4
Sivakanthan Kasinathan, Steven Henikoff
Animal and plant centromeres are embedded in repetitive "satellite" DNA, but are thought to be epigenetically specified. To define genetic characteristics of centromeres, we surveyed satellite DNA from diverse eukaryotes and identified variation in <10-bp dyad symmetries predicted to adopt non-B-form conformations. Organisms lacking centromeric dyad symmetries had binding sites for sequence-specific DNA-binding proteins with DNA-bending activity. For example, human and mouse centromeres are depleted for dyad symmetries, but are enriched for non-B-form DNA and are associated with binding sites for the conserved DNA-binding protein CENP-B, which is required for artificial centromere function but is paradoxically nonessential...
April 1, 2017: Molecular Biology and Evolution
https://www.readbyqxmd.com/read/29305387/simple-and-complex-centromeric-satellites-in-drosophila-sibling-species
#5
Paul B Talbert, Sivakanthan Kasinathan, Steven Henikoff
Centromeres are the chromosomal sites of assembly for kinetochores, the protein complexes that attach to spindle fibers and mediate separation of chromosomes to daughter cells in mitosis and meiosis. In most multicellular organisms, centromeres comprise a single specific family of tandem repeats-often 100-400 bp in length-found on every chromosome, typically in one location within heterochromatin. Drosophila melanogaster is unusual in that the heterochromatin contains many families of mostly short (5-12 bp) tandem repeats, none of which appear to be present at all centromeres, and none of which are found only at centromeres...
March 2018: Genetics
https://www.readbyqxmd.com/read/29225036/transcription-and-remodeling-produce-asymmetrically-unwrapped-nucleosomal-intermediates
#6
Srinivas Ramachandran, Kami Ahmad, Steven Henikoff
Nucleosomes are disrupted during transcription and other active processes, but the structural intermediates during nucleosome disruption in vivo are unknown. To identify intermediates, we mapped subnucleosomal protections in Drosophila cells using Micrococcal Nuclease followed by sequencing. At the first nucleosome position downstream of the transcription start site, we identified unwrapped intermediates, including hexasomes that lack either proximal or distal contacts. Inhibiting topoisomerases or depleting histone chaperones increased unwrapping, whereas inhibiting release of paused RNAPII or reducing RNAPII elongation decreased unwrapping...
December 21, 2017: Molecular Cell
https://www.readbyqxmd.com/read/29196559/remarkable-evolutionary-plasticity-of-centromeric-chromatin
#7
Steven Henikoff, Jitendra Thakur, Sivakanthan Kasinathan, Paul B Talbert
Centromeres were familiar to cell biologists in the late 19th century, but for most eukaryotes the basis for centromere specification has remained enigmatic. Much attention has been focused on the cenH3 (CENP-A) histone variant, which forms the foundation of the centromere. To investigate the DNA sequence requirements for centromere specification, we applied a variety of epigenomic approaches, which have revealed surprising diversity in centromeric chromatin properties. Whereas each point centromere of budding yeast is occupied by a single precisely positioned tetrameric nucleosome with one cenH3 molecule, the "regional" centromeres of fission yeast contain unphased presumably octameric nucleosomes with two cenH3s...
December 1, 2017: Cold Spring Harbor Symposia on Quantitative Biology
https://www.readbyqxmd.com/read/29065294/structural-biology-probing-the-origins-of-chromatin
#8
Steven Henikoff
A new study finds that archaeal histone dimers can multimerize into extended superhelical structures that mediate gene expression changes, providing possible insights into the transition to eukaryotic nucleosomes.
October 23, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28580964/corrigendum-chec-seq-kinetics-discriminates-transcription-factor-binding-sites-by-dna-sequence-and-shape-in-vivo
#9
Gabriel E Zentner, Sivakanthan Kasinathan, Beibei Xin, Remo Rohs, Steven Henikoff
This corrects the article DOI: 10.1038/ncomms15643.
June 5, 2017: Nature Communications
https://www.readbyqxmd.com/read/28580953/correspondence-reply-to-dna-shape-is-insufficient-to-explain-binding
#10
Sivakanthan Kasinathan, Gabriel E Zentner, Beibei Xin, Remo Rohs, Steven Henikoff
No abstract text is available yet for this article.
June 5, 2017: Nature Communications
https://www.readbyqxmd.com/read/28133760/capitalizing-on-disaster-establishing-chromatin-specificity-behind-the-replication-fork
#11
REVIEW
Srinivas Ramachandran, Kami Ahmad, Steven Henikoff
Eukaryotic genomes are packaged into nucleosomal chromatin, and genomic activity requires the precise localization of transcription factors, histone modifications and nucleosomes. Classic work described the progressive reassembly and maturation of bulk chromatin behind replication forks. More recent proteomics has detailed the molecular machines that accompany the replicative polymerase to promote rapid histone deposition onto the newly replicated DNA. However, localized chromatin features are transiently obliterated by DNA replication every S phase of the cell cycle...
April 2017: BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology
https://www.readbyqxmd.com/read/28079019/an-efficient-targeted-nuclease-strategy-for-high-resolution-mapping-of-dna-binding-sites
#12
Peter J Skene, Steven Henikoff
We describe Cleavage Under Targets and Release Using Nuclease (CUT&RUN), a chromatin profiling strategy in which antibody-targeted controlled cleavage by micrococcal nuclease releases specific protein-DNA complexes into the supernatant for paired-end DNA sequencing. Unlike Chromatin Immunoprecipitation (ChIP), which fragments and solubilizes total chromatin, CUT&RUN is performed in situ, allowing for both quantitative high-resolution chromatin mapping and probing of the local chromatin environment. When applied to yeast and human nuclei, CUT&RUN yielded precise transcription factor profiles while avoiding crosslinking and solubilization issues...
January 16, 2017: ELife
https://www.readbyqxmd.com/read/27924075/histone-variants-on-the-move-substrates-for-chromatin-dynamics
#13
REVIEW
Paul B Talbert, Steven Henikoff
Most histones are assembled into nucleosomes behind the replication fork to package newly synthesized DNA. By contrast, histone variants, which are encoded by separate genes, are typically incorporated throughout the cell cycle. Histone variants can profoundly change chromatin properties, which in turn affect DNA replication and repair, transcription, and chromosome packaging and segregation. Recent advances in the study of histone replacement have elucidated the dynamic processes by which particular histone variants become substrates of histone chaperones, ATP-dependent chromatin remodellers and histone-modifying enzymes...
February 2017: Nature Reviews. Molecular Cell Biology
https://www.readbyqxmd.com/read/27797823/mediator-binding-to-uass-is-broadly-uncoupled-from-transcription-and-cooperative-with-tfiid-recruitment-to-promoters
#14
Sebastian Grünberg, Steven Henikoff, Steven Hahn, Gabriel E Zentner
Mediator is a conserved, essential transcriptional coactivator complex, but its in vivo functions have remained unclear due to conflicting data regarding its genome-wide binding pattern obtained by genome-wide ChIP Here, we used ChEC-seq, a method orthogonal to ChIP, to generate a high-resolution map of Mediator binding to the yeast genome. We find that Mediator associates with upstream activating sequences (UASs) rather than the core promoter or gene body under all conditions tested. Mediator occupancy is surprisingly correlated with transcription levels at only a small fraction of genes...
November 15, 2016: EMBO Journal
https://www.readbyqxmd.com/read/27503998/mechanisms-of-nucleosome-dynamics-in-vivo
#15
REVIEW
Steven Henikoff
Nucleosomes function to tightly package DNA into chromosomes, but the nucleosomal landscape becomes disrupted during active processes such as replication, transcription, and repair. The realization that many proteins responsible for chromatin regulation are frequently mutated in cancer has drawn attention to chromatin dynamics; however, the basic mechanisms whereby nucleosomes are disrupted and reassembled is incompletely understood. Here, I present an overview of chromatin dynamics as has been elucidated in model organisms, in which our understanding is most advanced...
September 1, 2016: Cold Spring Harbor Perspectives in Medicine
https://www.readbyqxmd.com/read/27458904/epigenetics-cellular-memory-and-gene-regulation
#16
Steven Henikoff, John M Greally
The field described as 'epigenetics' has captured the imagination of scientists and the lay public. Advances in our understanding of chromatin and gene regulatory mechanisms have had impact on drug development, fueling excitement in the lay public about the prospects of applying this knowledge to address health issues. However, when describing these scientific advances as 'epigenetic', we encounter the problem that this term means different things to different people, starting within the scientific community and amplified in the popular press...
July 25, 2016: Current Biology: CB
https://www.readbyqxmd.com/read/27384170/cenpt-bridges-adjacent-cenpa-nucleosomes-on-young-human-%C3%AE-satellite-dimers
#17
Jitendra Thakur, Steven Henikoff
Nucleosomes containing the CenH3 (CENPA or CENP-A) histone variant replace H3 nucleosomes at centromeres to provide a foundation for kinetochore assembly. CENPA nucleosomes are part of the constitutive centromere associated network (CCAN) that forms the inner kinetochore on which outer kinetochore proteins assemble. Two components of the CCAN, CENPC and the histone-fold protein CENPT, provide independent connections from the ∼171-bp centromeric α-satellite repeat units to the outer kinetochore. However, the spatial relationship between CENPA nucleosomes and these two branches remains unclear...
September 2016: Genome Research
https://www.readbyqxmd.com/read/27062929/transcriptional-regulators-compete-with-nucleosomes-post-replication
#18
Srinivas Ramachandran, Steven Henikoff
Every nucleosome across the genome must be disrupted and reformed when the replication fork passes, but how chromatin organization is re-established following replication is unknown. To address this problem, we have developed Mapping In vivo Nascent Chromatin with EdU and sequencing (MINCE-seq) to characterize the genome-wide location of nucleosomes and other chromatin proteins behind replication forks at high temporal and spatial resolution. We find that the characteristic chromatin landscape at Drosophila promoters and enhancers is lost upon replication...
April 21, 2016: Cell
https://www.readbyqxmd.com/read/26933790/nucleosome-dynamics-during-chromatin-remodeling-in-vivo
#19
Srinivas Ramachandran, Steven Henikoff
Precise positioning of nucleosomes around regulatory sites is achieved by the action of chromatin remodelers, which use the energy of ATP to slide, evict or change the composition of nucleosomes. Chromatin remodelers act to bind nucleosomes, disrupt histone-DNA interactions and translocate the DNA around the histone core to reposition nucleosomes. Hence, remodeling is expected to involve nucleosomal intermediates with a structural organization that is distinct from intact nucleosomes. We describe the identification of a partially unwrapped nucleosome structure using methods that map histone-DNA contacts genome-wide...
2016: Nucleus
https://www.readbyqxmd.com/read/26877204/evolutionary-turnover-of-kinetochore-proteins-a-ship-of-theseus
#20
REVIEW
Ines A Drinnenberg, Steven Henikoff, Harmit S Malik
The kinetochore is a multiprotein complex that mediates the attachment of a eukaryotic chromosome to the mitotic spindle. The protein composition of kinetochores is similar across species as divergent as yeast and human. However, recent findings have revealed an unexpected degree of compositional diversity in kinetochores. For example, kinetochore proteins that are essential in some species have been lost in others, whereas new kinetochore proteins have emerged in other lineages. Even in lineages with similar kinetochore composition, individual kinetochore proteins have functionally diverged to acquire either essential or redundant roles...
July 2016: Trends in Cell Biology
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