Centrifugal elutriation (counterstreaming centrifugation) of cells

Pretlow, Thomas G.; Pretlow, Theresa P.

doi:10.1007/bf02781351

Cited by 60 publications

(15 citation statements)

References 43 publications

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“…Alternatively, we confirmed the cell cycle regulation of histone O-GlcNAcylation by using counter-flow centrifugal elutriation, a physical method that does not require any arresting agents but rather fractionates cells based on their increasing size as they progress through the cell cycle (29). Fig.…”

Section: Association Of Histone O-glcnacylation With Active and Inactsupporting

confidence: 53%

Modification of Histones by Sugar β-N-Acetylglucosamine (GlcNAc) Occurs on Multiple Residues, Including Histone H3 Serine 10, and Is Cell Cycle-regulated

Zhang

Roche

Nasheuer

et al. 2011

Journal of Biological Chemistry

123

112

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Section: Association Of Histone O-glcnacylation With Active and Inactsupporting

confidence: 53%

Modification of Histones by Sugar β-N-Acetylglucosamine (GlcNAc) Occurs on Multiple Residues, Including Histone H3 Serine 10, and Is Cell Cycle-regulated

Zhang

Roche

Nasheuer

et al. 2011

Journal of Biological Chemistry

123

112

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“…It is important to determine if synchronization protocol results in DNA damage, especially if the complexes being studied are normally involved in such mechanisms. In this scenario, it may be preferable to isolate cells at different phases of the cell cycle using flow cytometry [19] or centrifugal elutriation [20] instead. Cells are then trypsinized, washed once with 1× PBS, once with cold 1× PBS-T, and kept on ice until ready to proceed onto nuclear preparation and chromatin extraction.…”

Section: Methodsmentioning

confidence: 99%

Tracking Histone Variant Nucleosomes Across the Human Cell Cycle Using Biophysical, Biochemical, and Cytological Analyses

Walkiewicz

Bui

Quénet

et al. 2014

Methods in Molecular Biology

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Histone variants such as H3.3, macroH2A, H2A.Z, and CENP-A are important epigenetic modifiers of the chromatin state in eukaryotic genomes. The centromeric histone H3 variant CENP-A/CENH3 epigenetically marks centromeres and is required for assembly of the kinetochore complex, a region of the chromosome that is responsible for proper genome segregation during mitosis. Several diverse techniques using biochemical, cell biology, and biophysical approaches have been utilized to study the nature of the CENP-A nucleosome across the cell cycle. In this chapter, we describe methods for CENP-A nucleosome purification and separation of CENP-A from other core histones using traditional SDS-PAGE and more resolving techniques such as Triton acid urea (TAU) and two-dimensional gels. We also discuss methods for observation of CENP-A on chromatin fibers using immunofluorescence. Finally, we provide a detailed description of analysis of chromatin structures using atomic force microscopy.

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“…Centrifugal elutriation is a technique for the separation of cells by velocity sedimentation (Pretlow and Pretlow 1979). Asynchronous cells from an exponential culture in YPD were separated in different fractions.…”

Section: Methodsmentioning

confidence: 99%

The Spatiotemporal Program of Replication in the Genome of Lachancea kluyveri

Agier

Romano

Touzain

et al. 2013

Genome Biology and Evolution

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We generated a genome-wide replication profile in the genome of Lachancea kluyveri and assessed the relationship between replication and base composition. This species diverged from Saccharomyces cerevisiae before the ancestral whole genome duplication. The genome comprises eight chromosomes among which a chromosomal arm of 1 Mb has a G + C-content much higher than the rest of the genome. We identified 252 active replication origins in L. kluyveri and found considerable divergence in origin location with S. cerevisiae and with Lachancea waltii. Although some global features of S. cerevisiae replication are conserved: Centromeres replicate early, whereas telomeres replicate late, we found that replication origins both in L. kluyveri and L. waltii do not behave as evolutionary fragile sites. In L. kluyveri, replication timing along chromosomes alternates between regions of early and late activating origins, except for the 1 Mb GC-rich chromosomal arm. This chromosomal arm contains an origin consensus motif different from other chromosomes and is replicated early during S-phase. We showed that precocious replication results from the specific absence of late firing origins in this chromosomal arm. In addition, we found a correlation between GC-content and distance from replication origins as well as a lack of replication-associated compositional skew between leading and lagging strands specifically in this GC-rich chromosomal arm. These findings suggest that the unusual base composition in the genome of L. kluyveri could be linked to replication.

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Centrifugal elutriation (counterstreaming centrifugation) of cells

Cited by 60 publications

References 43 publications

Modification of Histones by Sugar β-N-Acetylglucosamine (GlcNAc) Occurs on Multiple Residues, Including Histone H3 Serine 10, and Is Cell Cycle-regulated

Modification of Histones by Sugar β-N-Acetylglucosamine (GlcNAc) Occurs on Multiple Residues, Including Histone H3 Serine 10, and Is Cell Cycle-regulated

Tracking Histone Variant Nucleosomes Across the Human Cell Cycle Using Biophysical, Biochemical, and Cytological Analyses

The Spatiotemporal Program of Replication in the Genome of Lachancea kluyveri

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