Single-molecule imaging of chromatin remodelers reveals role of ATPase in promoting fast kinetics of target search and dissociation from chromatin

eLife, Journal Year: 2021, Volume and Issue: 10

Published: July 27, 2021

Conserved ATP-dependent chromatin remodelers establish and maintain genome-wide architectures of regulatory DNA during cellular lifespan, but the temporal interactions between targets have been obscure. We performed live-cell single-molecule tracking for RSC, SWI/SNF, CHD1, ISW1, ISW2, INO80 remodeling complexes in budding yeast detected hyperkinetic behaviors chromatin-bound molecules that frequently transition to free state all complexes. Chromatin-bound display notably higher diffusion than nucleosomal histones, strikingly fast dissociation kinetics with 4–7 s mean residence times. These enhanced dynamics require ATP binding or hydrolysis by catalytic ATPase, uncovering an additional function its established role nucleosome remodeling. Kinetic simulations show multiple can repeatedly occupy same promoter region on a timescale minutes, implicating unending ‘tug-of-war’ controls temporally shifting window accessibility transcription initiation machinery.

Language: Английский

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Single-molecule tracking of transcription protein dynamics in living cells: seeing is believing, but what are we seeing?

Current Opinion in Genetics & Development, Journal Year: 2021, Volume and Issue: 67, P. 94 - 102

Published: Jan. 9, 2021

Language: Английский

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Quantitative imaging of transcription in living Drosophila embryos reveals the impact of core promoter motifs on promoter state dynamics

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: July 23, 2021

Abstract Genes are expressed in stochastic transcriptional bursts linked to alternating active and inactive promoter states. A major challenge transcription is understanding how composition dictates bursting, particularly multicellular organisms. We investigate two key Drosophila developmental motifs, the TATA box (TATA) Initiator (INR). Using live imaging embryos new computational methods, we demonstrate that bursting occurs on multiple timescales ranging from seconds minutes. TATA-containing promoters INR-containing exhibit distinct dynamics, with one or separate rate-limiting steps respectively. associated long states, high rates of polymerase initiation, short-lived, infrequent In contrast, INR motif leads which relates promoter-proximal pausing. Surprisingly, model suggests pausing not obligatory, but stochastically for a subset polymerases. Overall, our results provide rationale switching during zygotic genome activation.

Language: Английский

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Speed–Specificity Trade-Offs in the Transcription Factors Search for Their Genomic Binding Sites

Trends in Genetics, Journal Year: 2021, Volume and Issue: 37(5), P. 421 - 432

Published: Jan. 8, 2021

Language: Английский

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Transcription Factor Dynamics

Cold Spring Harbor Perspectives in Biology, Journal Year: 2021, Volume and Issue: 13(11), P. a040949 - a040949

Published: May 17, 2021

Feiyue Lu and Timothée Lionnet Institute for Systems Genetics Cell Biology Department, NYU School of Medicine, New York, York 10016, USA Correspondence: Timothee.lionnet{at}nyulangone.org

Language: Английский

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