Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells

Annamária Kakuk, Elza Friedländer, G. Vereb, Anita Kása, A. Balla, Tamás Balla, Ludwig M G Heilmeyer, P. Gergely, György Vereb

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Background: Previous immunohistochemical investigations could not detect PI4K230, an isoform of mammalian phosphatidylinositol 4-kinases (also called type III α), in the nucleus and nucleolus of cells in spite of its predicted nuclear localization signals. Methods: Immunofluorescent detection of PI4K230 and other PI4K isoforms was performed on formaldehyde (PFA) or ethanol fixed cells and rat brain cryosections. Costaining with nucleolin and the effect of siRNA, Triton X-100, DNase, and RNase treatments were also tested to determine the localization of PI4K230. Results: PI4K230 gives a prominent signal in the nucleolus of ethanol fixed rat brain cryosections and of several cell types in addition to its presence in the nucleus and cytoplasm. The PI4K230 immunoreactivity of the nucleolus is masked in PFA fixed cells, but it can be restored by treatment of PFA fixed cells with hot wet citrate buffer or by washing the cryosections with PBS prior to PFA fixation. Nucleolar PI4K230 occurs in a Triton X-100 resistant complex. Treatment of COS-7 cells with siRNA targeting PI4K230 and permeabilized B50 cells with DNase or RNase results in the loss of PI4K230 signal from the nucleolus. Conclusion: These experiments suggest the participation of PI4K230 in a DNase and RNase sensitive complex with a unique localization and function in the nucleolus.

Original languageEnglish
Pages (from-to)1174-1183
Number of pages10
JournalCytometry Part A
Volume69
Issue number12
DOIs
Publication statusPublished - Dec 1 2006

Fingerprint

1-Phosphatidylinositol 4-Kinase
Formaldehyde
Deoxyribonucleases
Ribonucleases
Octoxynol
Small Interfering RNA
Protein Isoforms
Ethanol
Cell Nucleolus
Nuclear Localization Signals
COS Cells
Brain
Cell Nucleus
Citric Acid
Buffers
Cytoplasm

Keywords

  • Cell lines HCN, B50, COS-7
  • Epitope masking
  • Immunofluorescence
  • Nucleolus
  • Phosphatidylinositol 4-kinase isoforms
  • PI4K230
  • Rat brain

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells. / Kakuk, Annamária; Friedländer, Elza; Vereb, G.; Kása, Anita; Balla, A.; Balla, Tamás; Heilmeyer, Ludwig M G; Gergely, P.; Vereb, György.

In: Cytometry Part A, Vol. 69, No. 12, 01.12.2006, p. 1174-1183.

Research output: Contribution to journalArticle

Kakuk, A, Friedländer, E, Vereb, G, Kása, A, Balla, A, Balla, T, Heilmeyer, LMG, Gergely, P & Vereb, G 2006, 'Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells', Cytometry Part A, vol. 69, no. 12, pp. 1174-1183. https://doi.org/10.1002/cyto.a.20347
Kakuk, Annamária ; Friedländer, Elza ; Vereb, G. ; Kása, Anita ; Balla, A. ; Balla, Tamás ; Heilmeyer, Ludwig M G ; Gergely, P. ; Vereb, György. / Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells. In: Cytometry Part A. 2006 ; Vol. 69, No. 12. pp. 1174-1183.
@article{641521f65a4c4ab5b6cbfe74a4d1cd74,
title = "Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells",
abstract = "Background: Previous immunohistochemical investigations could not detect PI4K230, an isoform of mammalian phosphatidylinositol 4-kinases (also called type III α), in the nucleus and nucleolus of cells in spite of its predicted nuclear localization signals. Methods: Immunofluorescent detection of PI4K230 and other PI4K isoforms was performed on formaldehyde (PFA) or ethanol fixed cells and rat brain cryosections. Costaining with nucleolin and the effect of siRNA, Triton X-100, DNase, and RNase treatments were also tested to determine the localization of PI4K230. Results: PI4K230 gives a prominent signal in the nucleolus of ethanol fixed rat brain cryosections and of several cell types in addition to its presence in the nucleus and cytoplasm. The PI4K230 immunoreactivity of the nucleolus is masked in PFA fixed cells, but it can be restored by treatment of PFA fixed cells with hot wet citrate buffer or by washing the cryosections with PBS prior to PFA fixation. Nucleolar PI4K230 occurs in a Triton X-100 resistant complex. Treatment of COS-7 cells with siRNA targeting PI4K230 and permeabilized B50 cells with DNase or RNase results in the loss of PI4K230 signal from the nucleolus. Conclusion: These experiments suggest the participation of PI4K230 in a DNase and RNase sensitive complex with a unique localization and function in the nucleolus.",
keywords = "Cell lines HCN, B50, COS-7, Epitope masking, Immunofluorescence, Nucleolus, Phosphatidylinositol 4-kinase isoforms, PI4K230, Rat brain",
author = "Annam{\'a}ria Kakuk and Elza Friedl{\"a}nder and G. Vereb and Anita K{\'a}sa and A. Balla and Tam{\'a}s Balla and Heilmeyer, {Ludwig M G} and P. Gergely and Gy{\"o}rgy Vereb",
year = "2006",
month = "12",
day = "1",
doi = "10.1002/cyto.a.20347",
language = "English",
volume = "69",
pages = "1174--1183",
journal = "Cytometry. Part A : the journal of the International Society for Analytical Cytology",
issn = "1552-4922",
publisher = "Wiley-Liss Inc.",
number = "12",

}

TY - JOUR

T1 - Nucleolar localization of phosphatidylinositol 4-kinase PI4K230 in various mammalian cells

AU - Kakuk, Annamária

AU - Friedländer, Elza

AU - Vereb, G.

AU - Kása, Anita

AU - Balla, A.

AU - Balla, Tamás

AU - Heilmeyer, Ludwig M G

AU - Gergely, P.

AU - Vereb, György

PY - 2006/12/1

Y1 - 2006/12/1

N2 - Background: Previous immunohistochemical investigations could not detect PI4K230, an isoform of mammalian phosphatidylinositol 4-kinases (also called type III α), in the nucleus and nucleolus of cells in spite of its predicted nuclear localization signals. Methods: Immunofluorescent detection of PI4K230 and other PI4K isoforms was performed on formaldehyde (PFA) or ethanol fixed cells and rat brain cryosections. Costaining with nucleolin and the effect of siRNA, Triton X-100, DNase, and RNase treatments were also tested to determine the localization of PI4K230. Results: PI4K230 gives a prominent signal in the nucleolus of ethanol fixed rat brain cryosections and of several cell types in addition to its presence in the nucleus and cytoplasm. The PI4K230 immunoreactivity of the nucleolus is masked in PFA fixed cells, but it can be restored by treatment of PFA fixed cells with hot wet citrate buffer or by washing the cryosections with PBS prior to PFA fixation. Nucleolar PI4K230 occurs in a Triton X-100 resistant complex. Treatment of COS-7 cells with siRNA targeting PI4K230 and permeabilized B50 cells with DNase or RNase results in the loss of PI4K230 signal from the nucleolus. Conclusion: These experiments suggest the participation of PI4K230 in a DNase and RNase sensitive complex with a unique localization and function in the nucleolus.

AB - Background: Previous immunohistochemical investigations could not detect PI4K230, an isoform of mammalian phosphatidylinositol 4-kinases (also called type III α), in the nucleus and nucleolus of cells in spite of its predicted nuclear localization signals. Methods: Immunofluorescent detection of PI4K230 and other PI4K isoforms was performed on formaldehyde (PFA) or ethanol fixed cells and rat brain cryosections. Costaining with nucleolin and the effect of siRNA, Triton X-100, DNase, and RNase treatments were also tested to determine the localization of PI4K230. Results: PI4K230 gives a prominent signal in the nucleolus of ethanol fixed rat brain cryosections and of several cell types in addition to its presence in the nucleus and cytoplasm. The PI4K230 immunoreactivity of the nucleolus is masked in PFA fixed cells, but it can be restored by treatment of PFA fixed cells with hot wet citrate buffer or by washing the cryosections with PBS prior to PFA fixation. Nucleolar PI4K230 occurs in a Triton X-100 resistant complex. Treatment of COS-7 cells with siRNA targeting PI4K230 and permeabilized B50 cells with DNase or RNase results in the loss of PI4K230 signal from the nucleolus. Conclusion: These experiments suggest the participation of PI4K230 in a DNase and RNase sensitive complex with a unique localization and function in the nucleolus.

KW - Cell lines HCN, B50, COS-7

KW - Epitope masking

KW - Immunofluorescence

KW - Nucleolus

KW - Phosphatidylinositol 4-kinase isoforms

KW - PI4K230

KW - Rat brain

UR - http://www.scopus.com/inward/record.url?scp=33845954519&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845954519&partnerID=8YFLogxK

U2 - 10.1002/cyto.a.20347

DO - 10.1002/cyto.a.20347

M3 - Article

C2 - 17131383

AN - SCOPUS:33845954519

VL - 69

SP - 1174

EP - 1183

JO - Cytometry. Part A : the journal of the International Society for Analytical Cytology

JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology

SN - 1552-4922

IS - 12

ER -