Visualizing the metazoan proliferation-quiescence decision in vivo

Adikes, Rebecca C.; Kohrman, Abraham; Martinez, Michael A Q; Palmisano, Nicholas J.; Smith, Jayson J.; Medwig-Kinney, Taylor N.; Min, Mingwei; Sallee, Maria D.; Ahmed, Ononnah B; Kim, Nuri; Liu, Simeiyun; Morabito, Robert D; Weeks, Nicholas; Zhao, Qinyun; Zhang, Wan; Feldman, Jessica L.; Barkoulas, Michalis; Pani, Ariel M.; Spencer, Steven J.; Martin, Benjamin L.; Matus, David Q.

doi:10.7554/elife.63265

Cited by 43 publications

(72 citation statements)

References 99 publications

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“…Interestingly, 28% (12/43) of animals treated with swsn-4(RNAi) were absent of SMs on either the left or the right side, whereas 100% (30/30) control animals had SMs on both sides, which may indicate a defect in specification, early cell division, and/or migration of SMs. To quantify cell cycle state, we measured localization of an SM-specific CDK sensor, which uses a fragment of mammalian DNA Helicase B (DHB) fused to two copies of mKate2 (36,70). In cells with low CDK activity that are quiescent or post-mitotic, the ratiometric CDK sensor is strongly nuclear localized (36,68,70).…”

Section: Resultsmentioning

confidence: 99%

“…We generated two anti-GFP nanobody constructs, using conserved cis-regulatory elements from the cdh-3 and egl-43 promoters (22,24,39,67,68) and introduced them into a strain containing the endogenous GFPtagged allele of swsn-4 as well as background AC and BM reporters (Fig 3 B-C). The cdh-3-driven nanobody transgene (cdh-3p::antiGFP-nanobody::p2a::his-58::mCherry) resulted in a weak reduction of GFP::SWSN-4 levels with no significant difference in fluorescence expression in the AC compared to wildtype animals (6% depletion; n=80 (36,70). In cells with low CDK activity that are quiescent or post-mitotic, the ratiometric CDK sensor is strongly nuclear localized (36,68,70).…”

Section: The Swi/snf Atpase Swsn-4 Provides Dose-dependent Regulation Of Ac Invasionmentioning

confidence: 99%

“…The cdh-3-driven nanobody transgene (cdh-3p::antiGFP-nanobody::p2a::his-58::mCherry) resulted in a weak reduction of GFP::SWSN-4 levels with no significant difference in fluorescence expression in the AC compared to wildtype animals (6% depletion; n=80 (36,70). In cells with low CDK activity that are quiescent or post-mitotic, the ratiometric CDK sensor is strongly nuclear localized (36,68,70). In cycling cells with increasing CDK activity, the CDK sensor progressively translocates from the nucleus to the cytosoplasm, with a ratio approaching 1.0 in S phase and >1 in cells in G2 (36).…”

Section: The Swi/snf Atpase Swsn-4 Provides Dose-dependent Regulation Of Ac Invasionmentioning

confidence: 99%

“…In cells with low CDK activity that are quiescent or post-mitotic, the ratiometric CDK sensor is strongly nuclear localized (36,68,70). In cycling cells with increasing CDK activity, the CDK sensor progressively translocates from the nucleus to the cytosoplasm, with a ratio approaching 1.0 in S phase and >1 in cells in G2 (36). Thus, the cytoplasmic: suggesting that the wild-type AC exists in a CDK low G0, pro-invasive state (Fig 4 A, E-F).…”

Section: The Swi/snf Atpase Swsn-4 Provides Dose-dependent Regulation Of Ac Invasionmentioning

confidence: 99%

“…This finding parallels similar studies in cancer (31–34) and C. elegans mesoblast development (35). Examination using a CDK activity sensor (36) revealed assembly-specific contributions to AC invasion: whereas BAF promotes AC invasion in a cell cycle-dependent manner, PBAF contributes to invasion in a cell cycle-independent manner. Finally, we utilized the auxin-inducible degron (AID) system combined with PBAF RNAi to achieve strong PBAF subunit depletion in the AC, which resulted in loss of both AC invasion and adhesion to the BM.…”

Section: Introductionmentioning

confidence: 99%

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The SWI/SNF chromatin remodeling assemblies BAF and PBAF differentially regulate cell cycle exit and cellular invasionin vivo

Smith

Xiao

et al. 2021

Preprint

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Chromatin remodeling complexes, such as the SWItching defective/Sucrose Non-Fermenting (SWI/SNF) ATP-dependent chromatin remodeling complex, coordinate metazoan development through broad regulation of chromatin accessibility and transcription, ensuring normal cell cycle control and cellular differentiation in a lineage-specific and temporally restricted manner. Mutations in subunits of chromatin regulating factors are associated with a variety of diseases and cancer metastasis co-opts cellular invasion found in healthy cells during development. Here we utilize Caenorhabditis elegans anchor cell (AC) invasion as an in vivo model to identify the suite of chromatin and chromatin regulating factors (CRFs) that promote cellular invasiveness. We demonstrate that the SWI/SNF ATP-dependent chromatin remodeling complex is a critical regulator of AC invasion, with pleiotropic effects on both G0/G1 cell cycle arrest and activation of invasive machinery. Using targeted protein degradation and RNA interference (RNAi), we show that SWI/SNF contributes to AC invasion in a dose-dependent fashion, with lower levels of activity in the AC corresponding to aberrant cell cycle entry and increased loss of invasion. Finally, we implicate the SWI/SNF BAF assembly in the regulation of the cell cycle, whereas our data suggests that the SWI/SNF PBAF assembly promotes AC attachment to the basement membrane (BM) and promotes the activation of the invasive machinery. Together these findings demonstrate that the SWI/SNF complex is necessary for two essential components of AC invasion: arresting cell cycle progression and remodeling the BM. The work here provides valuable single-cell mechanistic insight into the contributions of SWI/SNF assembly and subunit-specific disruptions to tumorigenesis and cancer metastasis.

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Section: Resultsmentioning

confidence: 99%

Section: The Swi/snf Atpase Swsn-4 Provides Dose-dependent Regulation Of Ac Invasionmentioning

confidence: 99%

Section: The Swi/snf Atpase Swsn-4 Provides Dose-dependent Regulation Of Ac Invasionmentioning

confidence: 99%

Section: The Swi/snf Atpase Swsn-4 Provides Dose-dependent Regulation Of Ac Invasionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The SWI/SNF chromatin remodeling assemblies BAF and PBAF differentially regulate cell cycle exit and cellular invasionin vivo

Smith

Xiao

et al. 2021

Preprint

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Extreme Sensitivity of Fitness to Environmental Conditions: Lessons from #1BigBatch

et al. 2023

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The phrase “survival of the fittest” has become an iconic descriptor of how natural selection works. And yet, precisely measuring fitness, even for single-celled microbial populations growing in controlled laboratory conditions, remains a challenge. While numerous methods exist to perform these measurements, including recently developed methods utilizing DNA barcodes, all methods are limited in their precision to differentiate strains with small fitness differences. In this study, we rule out some major sources of imprecision, but still find that fitness measurements vary substantially from replicate to replicate. Our data suggest that very subtle and difficult to avoid environmental differences between replicates create systematic variation across fitness measurements. We conclude by discussing how fitness measurements should be interpreted given their extreme environment dependence. This work was inspired by the scientific community who followed us and gave us tips as we live tweeted a high-replicate fitness measurement experiment at #1BigBatch.

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