open access

Vol 53, No 2 (2015)
Original paper
Submitted: 2015-04-07
Accepted: 2015-07-20
Published online: 2015-07-21
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The interactions between SATB1 and F-actin are important for mechanisms of active cell death

Dariusz Grzanka, Anna E. Kowalczyk, Magdalena Izdebska, Anna Klimaszewska-Wisniewska, Maciej Gagat
DOI: 10.5603/fhc.a2015.0018
·
Pubmed: 26315726
·
Folia Histochem Cytobiol 2015;53(2):152-161.

open access

Vol 53, No 2 (2015)
ORIGINAL PAPERS
Submitted: 2015-04-07
Accepted: 2015-07-20
Published online: 2015-07-21

Abstract

Introduction. The direct involvement of nuclear actin filaments in gene transcription and remodeling of chromatin is still debatable. However, nuclear localization of F-actin and its interactions with other nuclear matrix proteins have been reported. The aim of the study was to estimate the interactions between nuclear F-actin and one of the matrix proteins, special AT-rich sequence-binding protein 1 (SATB1), during active cell death induced in vitro by geldanamycin (GA).

Material and methods. The expression of SATB1 was modified by the transfection of non-aggressive breast cancer MCF-7 cells with siRNA against SATB1 or expression plasmid with cloned cDNA of SATB1. The amount and localization of F-actin were altered by changes of cofilin-1 (CFL1) expression in MCF-7 cells. The association between SATB1 and F-actin during GA-induced cell death was analyzed using confocal and transmission electron microscopy.

Results. Our studies revealed the colocalization between nuclear F-actin and SATB1 protein, during GA-induced death of breast cancer MCF-7 cells. The colocalization was enhanced in cells with overexpressed SATB1 and cofilin-1. At the ultrastructural level the SATB1 and F-actin complexes were seen at the border of condensed and decondensed chromatin. The presence of SATB1/F-actin molecular complexes was confirmed by magnetic separation of F-actin and interacting proteins.

Conclusion. We suggest that the molecular interactions between SATB1 and F-actin are necessary for active cell death to occur.

Abstract

Introduction. The direct involvement of nuclear actin filaments in gene transcription and remodeling of chromatin is still debatable. However, nuclear localization of F-actin and its interactions with other nuclear matrix proteins have been reported. The aim of the study was to estimate the interactions between nuclear F-actin and one of the matrix proteins, special AT-rich sequence-binding protein 1 (SATB1), during active cell death induced in vitro by geldanamycin (GA).

Material and methods. The expression of SATB1 was modified by the transfection of non-aggressive breast cancer MCF-7 cells with siRNA against SATB1 or expression plasmid with cloned cDNA of SATB1. The amount and localization of F-actin were altered by changes of cofilin-1 (CFL1) expression in MCF-7 cells. The association between SATB1 and F-actin during GA-induced cell death was analyzed using confocal and transmission electron microscopy.

Results. Our studies revealed the colocalization between nuclear F-actin and SATB1 protein, during GA-induced death of breast cancer MCF-7 cells. The colocalization was enhanced in cells with overexpressed SATB1 and cofilin-1. At the ultrastructural level the SATB1 and F-actin complexes were seen at the border of condensed and decondensed chromatin. The presence of SATB1/F-actin molecular complexes was confirmed by magnetic separation of F-actin and interacting proteins.

Conclusion. We suggest that the molecular interactions between SATB1 and F-actin are necessary for active cell death to occur.

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Keywords

SATB1; F-actin; cofilin-1; geldanamycin; apoptosis; MCF-7 cells; protein interactions; confocal microscopy; electron microscopy

About this article
Title

The interactions between SATB1 and F-actin are important for mechanisms of active cell death

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 53, No 2 (2015)

Article type

Original paper

Pages

152-161

Published online

2015-07-21

DOI

10.5603/fhc.a2015.0018

Pubmed

26315726

Bibliographic record

Folia Histochem Cytobiol 2015;53(2):152-161.

Keywords

SATB1
F-actin
cofilin-1
geldanamycin
apoptosis
MCF-7 cells
protein interactions
confocal microscopy
electron microscopy

Authors

Dariusz Grzanka
Anna E. Kowalczyk
Magdalena Izdebska
Anna Klimaszewska-Wisniewska
Maciej Gagat

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