Limited Proteolysis-Coupled Mass Spectrometry Identifies Phosphatidylinositol 4,5-Bisphosphate Effectors in Human Nuclear Proteome

Sztacho, Martin; Šalovská, Barbora; Červenka, Jakub; Balaban, Can; Hoboth, Peter; Hozák, Pavel

doi:https://dx.doi.org/10.3390/cells10010068

Number of the records: 1

Limited Proteolysis-Coupled Mass Spectrometry Identifies Phosphatidylinositol 4,5-Bisphosphate Effectors in Human Nuclear Proteome

1.

SYSNO ASEP	0541499
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Limited Proteolysis-Coupled Mass Spectrometry Identifies Phosphatidylinositol 4,5-Bisphosphate Effectors in Human Nuclear Proteome
Author(s)	Sztacho, Martin (UMG-J)_ORCID Šalovská, Barbora (UMG-J) Červenka, Jakub (UZFG-Y)_ORCID Balaban, Can (UMG-J) Hoboth, Peter (UMG-J)_ORCID Hozák, Pavel (UMG-J)_{RID, ORCID}
Number of authors	6
Article number	68
Source Title	Cells. - : MDPI Roč. 10, č. 1 (2021)
Number of pages	16 s.
Publication form	Online - E
Language	eng - English
Country	CH - Switzerland
Keywords	nucleus ; limited proteolysis ; mass spectrometry ; phosphoinositides ; phosphatidylinositol 4 ; 5-bisphosphate
Subject RIV	EA - Cell Biology
OECD category	Cell biology
R&D Projects	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)
	LM2018129 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	EF16_013/0001775 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
Method of publishing	Open access
Institutional support	UMG-J - RVO:68378050 ; UZFG-Y - RVO:67985904
UT WOS	000610015300001
DOI	10.3390/cells10010068
Annotation	Specific nuclear sub-compartments that are regions of fundamental processes such as gene expression or DNA repair, contain phosphoinositides (PIPs). PIPs thus potentially represent signals for the localization of specific proteins into different nuclear functional domains. We performed limited proteolysis followed by label-free quantitative mass spectrometry and identified nuclear protein effectors of the most abundant PIP-phosphatidylinositol 4,5-bisphosphate (PIP2). We identified 515 proteins with PIP2-binding capacity of which 191 'exposed' proteins represent a direct PIP2 interactors and 324 'hidden' proteins, where PIP2 binding was increased upon trypsin treatment. Gene ontology analysis revealed that 'exposed' proteins are involved in the gene expression as regulators of Pol II, mRNA splicing, and cell cycle. They localize mainly to non-membrane bound organelles-nuclear speckles and nucleolus and are connected to the actin nucleoskeleton. 'Hidden' proteins are linked to the gene expression, RNA splicing and transport, cell cycle regulation, and response to heat or viral infection. These proteins localize to the nuclear envelope, nuclear pore complex, or chromatin. Bioinformatic analysis of peptides bound in both groups revealed that PIP2-binding motifs are in general hydrophilic. Our data provide an insight into the molecular mechanism of nuclear PIP2 protein interaction and advance the methodology applicable for further studies of PIPs or other protein ligands.
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.mdpi.com/2073-4409/10/1/68

Number of the records: 1