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Nanoscale mapping of nuclear phosphatidylinositol phosphate landscape by dual-color dSTORM
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SYSNO ASEP 0541498 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Nanoscale mapping of nuclear phosphatidylinositol phosphate landscape by dual-color dSTORM Author(s) Hoboth, Peter (UMG-J) ORCID
Sztacho, Martin (UMG-J) ORCID
Šebesta, O. (CZ)
Schätz, M. (CZ)
Castano, Enrique (UMG-J)
Hozák, Pavel (UMG-J) RID, ORCIDArticle number 158890 Source Title Biochimica Et Biophysica Acta-Molecular and Cell Biology of Lipids. - : Elsevier - ISSN 1388-1981
Roč. 1866, č. 5 (2021)Number of pages 16 s. Publication form Online - E Language eng - English Country GB - United Kingdom Keywords Cell nucleus ; Nuclear speckles ; Nucleolus ; Phosphatidylinositol phosphates ; RNA polymerase II ; STORM Subject RIV EA - Cell Biology OECD category Cell biology R&D Projects ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LM2018129 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) EF16_013/0001775 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA19-05608S GA ČR - Czech Science Foundation (CSF) GA18-19714S GA ČR - Czech Science Foundation (CSF) LTC19048 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LTC20024 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Limited access Institutional support UMG-J - RVO:68378050 UT WOS 000636044300002 DOI 10.1016/j.bbalip.2021.158890 Annotation Current models of gene expression, which are based on single-molecule localization microscopy, acknowledge protein clustering and the formation of transcriptional condensates as a driving force of gene expression. However, these models largely omit the role of nuclear lipids and amongst them nuclear phosphatidylinositol phosphates (PIPs) in particular. Moreover, the precise distribution of nuclear PIPs in the functional sub-nuclear domains remains elusive. The direct stochastic optical reconstruction microscopy (dSTORM) provides an unprecedented resolution in biological imaging. Therefore, its use for imaging in the densely crowded cell nucleus is desired but also challenging. Here we present a dual-color dSTORM imaging and image analysis of nuclear PI(4,5)P2, PI(3,4)P2 and PI(4)P distribution while preserving the context of nuclear architecture. In the nucleoplasm, PI(4,5)P2 and PI(3,4)P2 co-pattern in close proximity with the subset of RNA polymerase II foci. PI(4,5)P2 is surrounded by fibrillarin in the nucleoli and all three PIPs are dispersed within the matrix formed by the nuclear speckle protein SON. PI(4,5)P2 is the most abundant nuclear PIP, while PI(4)P is a precursor for the biosynthesis of PI(4,5)P2 and PI(3,4)P2. Therefore, our data are relevant for the understanding the roles of nuclear PIPs and provide further evidence for the model in which nuclear PIPs represent a localization signal for the formation of lipo-ribonucleoprotein hubs in the nucleus. The discussed experimental pipeline is applicable for further functional studies on the role of other nuclear PIPs in the regulation of gene expression and beyond. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2021 Electronic address https://www.sciencedirect.com/science/article/pii/S1388198121000160?via%3Dihub
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