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001-es BibID:	BIBFORM060655
Első szerző:	Szalóki Nikoletta (biológus)
Cím:	Evidence for Homodimerization of the c-Fos Transcription Factor in Live Cells Revealed by Fluorescence Microscopy and Computer Modeling / Nikoletta Szalóki, Jan Wolfgang Krieger, István Komáromi, Katalin Tóth, György Vámosi
Dátum:	2015
ISSN:	0270-7306
Megjegyzések:	The c-Fos and c-Jun transcription factors, members of the activator protein 1 (AP-1) complex, form heterodimers and bind to DNA via a basic leucine zipper and regulate the cell cycle, apoptosis, differentiation, etc. Purified c-Jun leucine zipper fragments could also form stable homodimers, whereas c-Fos leucine zipper homodimers were found to be much less stable in earlier in vitro studies. The importance of c-Fos overexpression in tumors and the controversy in the literature concerning c-Fos homodimerization prompted us to investigate Fos homodimerization. Förster resonance energy transfer (FRET) and molecular brightness analysis of fluorescence correlation spectroscopy data from live HeLa cells transfected with fluorescent-protein-tagged c-Fos indicated that c-Fos formed homodimers. We developed a method to determine the absolute concentrations of transfected and endogenous c-Fos and c-Jun, which allowed us to determine dissociation constants of c-Fos homodimers (Kd = 6.7 ? 1.7 ?M) and c-Fos?c-Jun heterodimers (on the order of 10 to 100 nM) from FRET titrations. Imaging fluorescence cross-correlation spectroscopy (SPIM-FCCS) and molecular dynamics modeling confirmed that c-Fos homodimers were stably associated and could bind to the chromatin. Our results establish c-Fos homodimers as a novel form of the AP-1 complex that may be an autonomous transcription factor in c-Fos-overexpressing tissues and could contribute to tumor development.
Tárgyszavak:	Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban c-Fos homodimerization dissociation constant fluorescence correlation spectroscopy SPIM-FCCS FRET live cell imaging oncogene protein-protein interactions protein-DNA interactions
Megjelenés:	Molecular And Cellular Biology. - 35 : 21 (2015), p. 3785-3798. -
További szerzők:	Krieger, Jan Wolfgang Komáromi István (1957-) (vegyész, molekuláris biológus, biokémikus) Tóth Katalin (Heidelberg) Vámosi György (1967-) (biofizikus)
Pályázati támogatás:	K103965 OTKA TÁMOP-4.2.2.A-11/1/KONV-2012-0023 TÁMOP MÖB/21-1/2013 Egyéb TÁMOP-4.2.4.A/2- 11/1-2012-0001 TÁMOP
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