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Detecting chromatin interactions along and between sister chromatids with SisterC

Marlies E. Oomen, Adam K. Hedger, Jonathan K. Watts, Job Dekker
doi: https://doi.org/10.1101/2020.03.10.986208
Marlies E. Oomen
1Program in Systems Biology, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA
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Adam K. Hedger
2RNA Therapeutics Institute and Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA
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Jonathan K. Watts
2RNA Therapeutics Institute and Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA
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Job Dekker
1Program in Systems Biology, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA
3Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA
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  • For correspondence: job.dekker@umassmed.edu
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Abstract

Accurate chromosome segregation requires chromosome compaction with concordant disentanglement of the two sister chromatids. This process has been studied extensively by microscopy but has remained a challenge for genomic methods, such as Hi-C, because sister chromatids have identical DNA sequences. Here we describe SisterC, a chromosome conformation capture assay that can distinguish interactions between and within sister chromatids. The assay is based on BrdU incorporation during S-phase, which labels the newly replicated strands of the sister chromatids. This is followed by Hi-C, e.g. during different stages of mitosis, and the selective destruction of BrdU containing strands by UV/Hoechst treatment. After PCR amplification and sequencing of the remaining intact strands, this allows for the assignment of Hi-C products as inter- and intra-sister interactions by read orientation. We performed SisterC on mitotically arrested S. cerevisiae cells. As expected, we find prominent interactions and alignment of sister chromatids at their centromeres. Along the arms, sister chromatids are less precisely aligned with inter-sister connections every ~35kb. In many instances, inter-sister interactions do not involve the interaction of two identical loci but occur between cohesin binding sites that can be offset by 5 to 25kb. Along sister chromatids, extruding cohesin forms loops up to 50kb. Combined, SisterC allows the observation of the complex interplay between sister chromatid compaction and sister chromatid segregation as the cell transitions from late S-phase to mitosis. SisterC should be applicable to study mitotic events in a wide range of organisms and cell types.

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Posted March 11, 2020.
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Detecting chromatin interactions along and between sister chromatids with SisterC
Marlies E. Oomen, Adam K. Hedger, Jonathan K. Watts, Job Dekker
bioRxiv 2020.03.10.986208; doi: https://doi.org/10.1101/2020.03.10.986208
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Detecting chromatin interactions along and between sister chromatids with SisterC
Marlies E. Oomen, Adam K. Hedger, Jonathan K. Watts, Job Dekker
bioRxiv 2020.03.10.986208; doi: https://doi.org/10.1101/2020.03.10.986208

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