Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation
Abstract Fluorescent proteins (FP) are frequently used for studying proteins inside cells. In advanced fluorescence microscopy, FPs can report on additional intracellular variables. One variable is the local density near FPs, which can be useful in studying densities within cellular bio-condensates....
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-08-01
|
| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-40647-6 |
| _version_ | 1797558083777462272 |
|---|---|
| author | Khalil Joron Juliane Oliveira Viegas Liam Haas-Neill Sariel Bier Paz Drori Shani Dvir Patrick Siang Lin Lim Sarah Rauscher Eran Meshorer Eitan Lerner |
| author_facet | Khalil Joron Juliane Oliveira Viegas Liam Haas-Neill Sariel Bier Paz Drori Shani Dvir Patrick Siang Lin Lim Sarah Rauscher Eran Meshorer Eitan Lerner |
| author_sort | Khalil Joron |
| collection | DOAJ |
| description | Abstract Fluorescent proteins (FP) are frequently used for studying proteins inside cells. In advanced fluorescence microscopy, FPs can report on additional intracellular variables. One variable is the local density near FPs, which can be useful in studying densities within cellular bio-condensates. Here, we show that a reduction in fluorescence lifetimes of common monomeric FPs reports increased levels of local densities. We demonstrate the use of this fluorescence-based variable to report the distribution of local densities within heterochromatin protein 1α (HP1α) in mouse embryonic stem cells (ESCs), before and after early differentiation. We find that local densities within HP1α condensates in pluripotent ESCs are heterogeneous and cannot be explained by a single liquid phase. Early differentiation, however, induces a change towards a more homogeneous distribution of local densities, which can be explained as a liquid-like phase. In conclusion, we provide a fluorescence-based method to report increased local densities and apply it to distinguish between homogeneous and heterogeneous local densities within bio-condensates. |
| first_indexed | 2024-03-10T17:26:28Z |
| format | Article |
| id | doaj.art-87782aa354b54bb290ad2376c488131e |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:26:28Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-87782aa354b54bb290ad2376c488131e2023-11-20T10:10:39ZengNature PortfolioNature Communications2041-17232023-08-0114111810.1038/s41467-023-40647-6Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiationKhalil Joron0Juliane Oliveira Viegas1Liam Haas-Neill2Sariel Bier3Paz Drori4Shani Dvir5Patrick Siang Lin Lim6Sarah Rauscher7Eran Meshorer8Eitan Lerner9Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of JerusalemDepartment of Genetics, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat RamDepartment of Chemical and Physical Sciences, University of Toronto MississaugaDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of JerusalemDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of JerusalemDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of JerusalemDepartment of Genetics, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat RamDepartment of Chemical and Physical Sciences, University of Toronto MississaugaDepartment of Genetics, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Edmond J. Safra Campus, Givat RamDepartment of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of JerusalemAbstract Fluorescent proteins (FP) are frequently used for studying proteins inside cells. In advanced fluorescence microscopy, FPs can report on additional intracellular variables. One variable is the local density near FPs, which can be useful in studying densities within cellular bio-condensates. Here, we show that a reduction in fluorescence lifetimes of common monomeric FPs reports increased levels of local densities. We demonstrate the use of this fluorescence-based variable to report the distribution of local densities within heterochromatin protein 1α (HP1α) in mouse embryonic stem cells (ESCs), before and after early differentiation. We find that local densities within HP1α condensates in pluripotent ESCs are heterogeneous and cannot be explained by a single liquid phase. Early differentiation, however, induces a change towards a more homogeneous distribution of local densities, which can be explained as a liquid-like phase. In conclusion, we provide a fluorescence-based method to report increased local densities and apply it to distinguish between homogeneous and heterogeneous local densities within bio-condensates.https://doi.org/10.1038/s41467-023-40647-6 |
| spellingShingle | Khalil Joron Juliane Oliveira Viegas Liam Haas-Neill Sariel Bier Paz Drori Shani Dvir Patrick Siang Lin Lim Sarah Rauscher Eran Meshorer Eitan Lerner Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation Nature Communications |
| title | Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation |
| title_full | Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation |
| title_fullStr | Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation |
| title_full_unstemmed | Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation |
| title_short | Fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem-cell differentiation |
| title_sort | fluorescent protein lifetimes report densities and phases of nuclear condensates during embryonic stem cell differentiation |
| url | https://doi.org/10.1038/s41467-023-40647-6 |
| work_keys_str_mv | AT khaliljoron fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT julianeoliveiraviegas fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT liamhaasneill fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT sarielbier fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT pazdrori fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT shanidvir fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT patricksianglinlim fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT sarahrauscher fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT eranmeshorer fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation AT eitanlerner fluorescentproteinlifetimesreportdensitiesandphasesofnuclearcondensatesduringembryonicstemcelldifferentiation |