GPCR-induced calcium transients trigger nuclear actin assembly for chromatin dynamics

Wang, Y; Sherrard, Alice; Zhao, Bing; Melak, Michael; Trautwein, Jonathan; Kleinschnitz, Eva-Maria; Tsopoulidis, Nikolaos; Fackler, Oliver T.; Schwan, Carsten; Grosse, Robert

doi:10.1038/s41467-019-13322-y

Cited by 62 publications

(68 citation statements)

References 31 publications

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“…Importantly, FLIM-FRET also provides accurate quantification due to the independence of the fluorescence lifetime from the relative concentrations of the interacting proteins, and is independent of their diffusion rates (Llères et al, 2007). In conclusion, our assay measures close contacts in the nuclear space between distant nucleosomes, and thus provides a read-out of nanoscale chromatin compaction (Lou et al, 2019;Sobecki et al, 2016;Baarlink et al, 2017;Wang et al, 2019a).…”

Section: Nanoscale Chromatin Compaction Is Decreased In Set-2 Mutant mentioning

confidence: 71%

Cooperation between Caenorhabditis elegansCOMPASS and condensin in germline chromatin organization

Herbette

Robert

Bailly

et al. 2020

Preprint

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Deposition of histone H3 lysine 4 (H3K4) methylation at promoters by SET1/COMPASS is associated with context-dependent effects on gene expression and local changes in chromatin organization.Whether SET1/COMPASS also contributes to higher-order chromosome structure has not been investigated. Here, we address this question by quantitative FRET (Förster resonance energy transfer)based fluorescence lifetime imaging microscopy (FLIM) on C. elegans germ cells expressing histones H2B-eGFP and H2B-mCherry. We find that SET1/COMPASS subunits strongly influence meiotic chromosome organization, with marked effects on the close proximity between nucleosomes. We further show that inactivation of SET-2, the C. elegans homologue of SET1, or CFP-1, the chromatin targeting subunit of COMPASS, strongly enhance chromosome organization defects and loss of fertility resulting from depletion of condensin-II. Defects in chromosome morphology resulting from conditional inactivation of topoisomerase II, another structural component of chromosomes, were also aggravated in the absence of SET-2. Combined, our in vivo findings suggest a model in which the SET1/COMPASS histone methyltransferase complex plays a role in shaping meiotic chromosome in cooperation with the non-histone proteins condensin-II and topoisomerase.

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Section: Nanoscale Chromatin Compaction Is Decreased In Set-2 Mutant mentioning

confidence: 71%

Cooperation between Caenorhabditis elegansCOMPASS and condensin in germline chromatin organization

Herbette

Robert

Bailly

et al. 2020

Preprint

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“…To directly visualize nuclear actin filaments, we stably expressed nuclear actin chromobody (nAC) (Plessner et al, 2015) in the HaloTag-MVI HeLa cells and induced formation of transient actin filaments by 8 µM A23187 (Y. Wang et al, 2019) (Methods). Indeed, two color imaging of nAC-GFP and SiR-HaloTag-MVI revealed that MVI moved along nuclear actin filaments (Figure 3b, Supplementary Figures 17 and 18 and Supplementary Movies 6 and 7).…”

Section: Figure 3: Nuclear Myosin VI Moves On Nuclear Actin Filamentsmentioning

confidence: 99%

“…Nuclear actin filaments were visualized with a GFP tagged nuclear actin chromobody (Chromotek) and filaments were induced with 8 µM Ionphore A23187 (Sigma) (Y. Wang et al, 2019) prior to the measurements.…”

Section: Additional Cell Treatmentsmentioning

confidence: 99%

Myosin VI moves on nuclear actin filaments and supports long-range chromatin rearrangements

Große-Berkenbusch

Hettich

Kühn

et al. 2020

Preprint

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Nuclear myosin VI (MVI) enhances RNA polymerase IIdependent transcription, but the molecular mechanism is unclear. We used live cell single molecule tracking to follow individual MVI molecules inside the nucleus and observed micrometer-long motion of the motor. Besides static chromatin interactions lasting for tens of seconds, ATPase-dependent directed motion occurred with a velocity of 2 µm/s. The movement was frequently interrupted by short periods of slow restricted diffusion and increased in frequency upon stimulation of transcription. Mutagenesis and perturbation experiments demonstrated that nuclear MVI motion is independent of dimerization and occurs on nuclear actin filaments, which we also observed by two-color imaging. Using chromosome paint to quantify distances between chromosomes, we found that MVI is required for transcription-dependent longrange chromatin rearrangements. Our measurements reveal a transcription-coupled function of MVI in the nucleus, where it actively undergoes directed movement along nuclear actin filaments. Motion is potentially mediated by cooperating monomeric motors and might assist in enhancing transcription by supporting long-range chromatin rearrangements.

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“…Importantly, FRET is a phenomenon sensitive to fluorophore proximity that occurs efficiently only when 6 the donor and acceptor fluorescent fusion proteins are closely positioned (<10nm) in the 3D nuclear space following chromatin compaction. By measuring histone-histone proximity, this FRET assay can respond to changes in nucleosome spacing, and therefore provides a read-out of nanoscale chromatin compaction (38)(39)(40)(41). FLIM-FRET also provides accurate quantification due to the independence of the fluorescence lifetime from the relative concentrations of the interacting proteins, and is independent of their diffusion rates (42).…”

Section: Nanoscale Chromatin Compaction Is Decreased In Set-2 Mutant mentioning

confidence: 99%

Cooperation between Caenorhabditis elegans COMPASS and condensin in germline chromatin organization

Herbette

Robert

Bailly

et al. 2020

Preprint

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Background Histone-modifying activities play important roles in gene expression and influence higher-order genome organization. SET1/COMPASS (Complex Proteins Associated with Set1) deposits h istone H3 lysine 4 (H3K4) methylation at promoter regions and is associated with context-dependent effects on gene expression. Whether it also contributes to higher-order chromosome organization has not been explored. Results Using a quantitative FRET (Förster resonance energy transfer)-based fluorescence lifetime imaging microscopy (FLIM) approach to assay nanometer scale chromatin compaction in live animals, we reveal a novel role for SET1/COMPASS in structuring meiotic chromosomes in the C. elegans germline . Inactivation of SET-2, the C. elegans homologue of SET1, strongly enhanced chromosome organization defects and loss of fertility resulting from depletion of condensin-II, and aggravated defects in chromosome morphology resulting from inactivation of topoisomerase II, another major structural component of chromosomes. Loss of CFP-1, the chromatin targeting subunit of COMPASS, similarly affected germline chromatin compaction measured by FLIM-FRET and enhanced condensin-II knock-down phenotypes. Conclusions The data presented here are consistent with a role of SET1/ COMPASS in shaping meiotic chromosomes in the C. elegans germline. This new insight has important implications for how c hromatin-modifying complexes and histone modifications may cooperate with non histone-proteins to achieve proper chromosome organization, not only in meiosis, but also in mitosis.

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GPCR-induced calcium transients trigger nuclear actin assembly for chromatin dynamics

Cited by 62 publications

References 31 publications

Cooperation between Caenorhabditis elegansCOMPASS and condensin in germline chromatin organization

Cooperation between Caenorhabditis elegansCOMPASS and condensin in germline chromatin organization

Myosin VI moves on nuclear actin filaments and supports long-range chromatin rearrangements

Cooperation between Caenorhabditis elegans COMPASS and condensin in germline chromatin organization

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