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Nanoscale analysis of nuclear phosphatidylinositol phosphate distribution between nucleaoplasm and nuclear speckles

SYSNO ASEP	0555748
Document Type	O - Others
R&D Document Type	Others
Title	Nanoscale analysis of nuclear phosphatidylinositol phosphate distribution between nucleaoplasm and nuclear speckles
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	CZ - Czech Republic
Keywords	single-molecule localization microscopy ; nuclear phosphatidylinositol phosphates ; nucleoplasm ; RNA polymerase II transcription
Subject RIV	EB - Genetics ; Molecular Biology
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 microscopy (SMLM) provided an unprecedented insight into the sub-nuclear organization of proteins and nucleic acids. However, the role of the nuclear lipids in the establishment of the functional nuclear architecture, apart from the nuclear envelope, has been neglected. Nevertheless, the roles in gene expression of the nuclear lipids and phosphatidylinositol phosphates (PIPs) in particular started to emerge. Therefore, we implemented and optimized the SMLM-based approach for the quantitative evaluation of the nuclear PIP distribution while preserving the context of nuclear architecture. We have quantitatively characterized the spatial distribution of nuclear phosphatidylinositol 4,5- and 3,4-bisphosphate (PI(4,5)P2 and PI(3,4)P2, resp.) and showed that PI(4,5)P2 and PI(3,4)P2 localize within matrix of the nuclear speckle marker SON and in the nucleoplasmic foci. Moreover, we found PI(4,5)P2 and PI(3,4)P2 in the close proximity with the subset of RNAPII foci either in the nucleoplasm or at nuclear speckles. We started to investigate the cross-talk between nucleoplasmic and nuclear speckle-associated PI(4,5)P2 and PI(3,4)P2 pools and their possible roles in the regulation of RNAPII transcription. Our preliminary data suggest that upon transcription inhibition PI(4,5)P2 and PI(3,4)P2 accumulate within nuclear speckles. Therefore, nuclear speckles could play a role in the buffering of the nuclear PIP levels and thereby possibly regulate RNAPII transcription.
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