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Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions
Visualizing and measuring molecular-scale interactions in living cells represents a major challenge, but recent advances in microscopy are bringing us closer to achieving this goal. Single-molecule super-resolution microscopy enables high-resolution and sensitive imaging of the positions and movemen...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10028927/ https://www.ncbi.nlm.nih.gov/pubmed/36945633 http://dx.doi.org/10.1101/2023.03.08.531771 |
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author | Chen, Ziyuan Seman, Melissa Farhat, Ali Fyodorova, Yekaterina Biswas, Saikat Levashkevich, Alexander Freddolino, P. Lydia Biteen, Julie S. Ragunathan, Kaushik |
author_facet | Chen, Ziyuan Seman, Melissa Farhat, Ali Fyodorova, Yekaterina Biswas, Saikat Levashkevich, Alexander Freddolino, P. Lydia Biteen, Julie S. Ragunathan, Kaushik |
author_sort | Chen, Ziyuan |
collection | PubMed |
description | Visualizing and measuring molecular-scale interactions in living cells represents a major challenge, but recent advances in microscopy are bringing us closer to achieving this goal. Single-molecule super-resolution microscopy enables high-resolution and sensitive imaging of the positions and movement of molecules in living cells. HP1 proteins are important regulators of gene expression because they selectively bind and recognize H3K9 methylated (H3K9me) histones to form heterochromatin-associated protein complexes that silence gene expression. Here, we extended live-cell single-molecule tracking studies in fission yeast to determine how HP1 proteins interact with their binding partners in the nucleus. We measured how genetic perturbations that affect H3K9me alter the diffusive properties of HP1 proteins and each of their binding partners based on which we inferred their most likely interaction sites. Our results indicate that H3K9me promotes specific complex formation between HP1 proteins and their interactors in a spatially restricted manner, while attenuating their ability to form off-chromatin complexes. As opposed to being an inert platform or scaffold to direct HP1 binding, our studies propose a novel function for H3K9me as an active participant in enhancing HP1-associated complex formation in living cells. |
format | Online Article Text |
id | pubmed-10028927 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-100289272023-03-22 Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions Chen, Ziyuan Seman, Melissa Farhat, Ali Fyodorova, Yekaterina Biswas, Saikat Levashkevich, Alexander Freddolino, P. Lydia Biteen, Julie S. Ragunathan, Kaushik bioRxiv Article Visualizing and measuring molecular-scale interactions in living cells represents a major challenge, but recent advances in microscopy are bringing us closer to achieving this goal. Single-molecule super-resolution microscopy enables high-resolution and sensitive imaging of the positions and movement of molecules in living cells. HP1 proteins are important regulators of gene expression because they selectively bind and recognize H3K9 methylated (H3K9me) histones to form heterochromatin-associated protein complexes that silence gene expression. Here, we extended live-cell single-molecule tracking studies in fission yeast to determine how HP1 proteins interact with their binding partners in the nucleus. We measured how genetic perturbations that affect H3K9me alter the diffusive properties of HP1 proteins and each of their binding partners based on which we inferred their most likely interaction sites. Our results indicate that H3K9me promotes specific complex formation between HP1 proteins and their interactors in a spatially restricted manner, while attenuating their ability to form off-chromatin complexes. As opposed to being an inert platform or scaffold to direct HP1 binding, our studies propose a novel function for H3K9me as an active participant in enhancing HP1-associated complex formation in living cells. Cold Spring Harbor Laboratory 2023-10-19 /pmc/articles/PMC10028927/ /pubmed/36945633 http://dx.doi.org/10.1101/2023.03.08.531771 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Chen, Ziyuan Seman, Melissa Farhat, Ali Fyodorova, Yekaterina Biswas, Saikat Levashkevich, Alexander Freddolino, P. Lydia Biteen, Julie S. Ragunathan, Kaushik Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
title | Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
title_full | Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
title_fullStr | Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
title_full_unstemmed | Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
title_short | Tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
title_sort | tracking live-cell single-molecule dynamics enables measurements of heterochromatinassociated protein-protein interactions |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10028927/ https://www.ncbi.nlm.nih.gov/pubmed/36945633 http://dx.doi.org/10.1101/2023.03.08.531771 |
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