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Unraveling the phospholipid identity of the gene expression compartments by Q-DC-dSTORM

SYSNO ASEP	0555746
Document Type	O - Others
R&D Document Type	Others
Title	Unraveling the phospholipid identity of the gene expression compartments by Q-DC-dSTORM
Author(s)	Hoboth, Peter (UMG-J)_ORCID Sztacho, Martin (UMG-J)_ORCID Šebesta, O. (CZ) Hozák, Pavel (UMG-J)_{RID, ORCID}
Year of issue	2021
Language	eng - English
Country	DE - Germany
Keywords	quantitative dual-color direct stochastic optical reconstruction microscopy ; nuclear phosphatidylinositol phosphates ; nucleoplasm ; nuclear speckles ; RNA polymerase II transcription
Subject RIV	DB - Geology ; Mineralogy
OECD category	Cell biology
R&D Projects	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)
	ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Institutional support	UMG-J - RVO:68378050
Annotation	Single-molecule localization (SML) microscopy provided an unprecedented insight into the sub-nuclear organization of proteins and nucleic acids and helped to develop models that acknowledge formation of transcriptional condensates as a driving force of gene expression. However, the roles of nuclear lipids in the establishment of the functional nuclear architecture, apart from the nuclear envelope, has been neglected. Nevertheless, accumulating evidence suggests the involvement of nuclear lipids and particularly of phosphatidylinositol phosphates (PIPs) in gene expression. We used quantitative dual-color direct stochastic optical reconstruction microscopy (Q-DC-dSTORM) for the evaluation of the distribution of immunolabeled nuclear PIP while preserving the context of nuclear architecture. We revealed for the first time the spatial organization of nuclear phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), phosphatidylinositol 3,4-bisphosphate (PI(3,4)P2) and phosphatidylinositol 4-monophosphate (PI(4)P) within individual nuclear sub-compartments. 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. We used Q-DC-dSTORM and analyzed the spatial relationship of PI(4,5)P2, PI(3,4)P2 and PI(4)P with the nuclear speckle marker SON and with RNA polymerase II (RNAPII). We compared the real data with random SMLs which allowed us to assess if the co-patterning of the two probes is non-random and visualized NNDs using self-developed in cellulo color-coded maps [1,5]. We found all three PIPs dispersed within SON matrix in the nuclear speckles and PI(4,5)P2 and PI(3,4)P2 in the close proximity with the subset of RNAPII foci. We showed PI(4,5)P2, PI(3,4)P2 and PI(4)P within nuclear speckles and in the nucleoplasmic foci and PI(4,5)P2 and PI(3,4)P2 in the close proximity with the subset of RNAII foci either in the nucleoplasm or nuclear speckles.
Workplace	Institute of Molecular Genetics
Contact	Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217
Year of Publishing	2022

Number of the records: 1