Detailed Information on Publication Record
2007
Bioinformatic predictions and image analysis of localization and interactions of endonuclease G, AIF, and AMID in human cells
VAŘECHA, Miroslav, Jana AMRICHOVÁ, Michal ZIMMERMANN, Vladimír ULMAN, Pavel MATULA et. al.Basic information
Original name
Bioinformatic predictions and image analysis of localization and interactions of endonuclease G, AIF, and AMID in human cells
Name in Czech
Bioinformatická predikce a obrazová analýza lokalizace a interakcí endonukleázy G, AIF a AMIDu v lidských buňkách
Authors
VAŘECHA, Miroslav (203 Czech Republic, guarantor), Jana AMRICHOVÁ (203 Czech Republic), Michal ZIMMERMANN (203 Czech Republic), Vladimír ULMAN (203 Czech Republic), Pavel MATULA (203 Czech Republic) and Michal KOZUBEK (203 Czech Republic)
Edition
Book of Abstracts for 15th Euroconference on Apoptosis, 2007
Other information
Language
English
Type of outcome
Konferenční abstrakt
Field of Study
Genetics and molecular biology
Country of publisher
Slovenia
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
RIV identification code
RIV/00216224:14330/07:00020613
Organization unit
Faculty of Informatics
Keywords in English
apoptosis; cell death; autophagy; cancer
Tags
Tags
International impact, Reviewed
Změněno: 2/6/2010 11:53, Mgr. Miroslav Vařecha, Ph.D.
V originále
During apoptosis several mitochondrial proteins are released. They participate in caspase-independent nuclear DNA degradation, namely apoptosis-inducing factor (AIF, also PDCD8 or AIFM1) and endonuclease G. Another interesting protein, which is not located inside mitochondria, was expected to act similarly as AIF due to high sequence homology with AIF. This protein is AIF-homologous mitochondrion-associated inducer of death (AMID, also PRG3 or AIFM2). We studied the cellular localization and colocalization of proteins AIF, endonuclease G and AMID experimentally using designed mammalian expression vectors, which carry genes encoding the proteins of interest fused to the fluorescent proteins and using bioinformatic predictions, that analyze the amino acid sequence of the proteins with various algorithms. We also designed and applied the novel method of single-cell image analysis of the translocation of the fluorescent proteins into the nucleus. We confirmed the colocalization of AIF and endonuclease G in the mitochondria of human cells. We also analyzed their translocation from mitochondria to the nucleus during apoptosis. AMID was found to be cytoplasmic protein, bound to various cellular surfaces from their cytoplasmic side. Overexpression of fusion protein AMID-HcRed-tandem was not lethal to the cells or mitochondria. We did not observe its translocation into the nucleus during staurosporine-induced apoptosis. The possible role of AMID in apoptosis was not observed. Bioinformatic predictions and time-lapse FRET experiments were conducted to analyze the interactions of the studied proteins in living and fixed human cells. Our results contribute to the comprehension of localization, interactions and functions of AMID, AIF, and endonuclease G in human cells.
In Czech
Během apoptózy se uvolní několik protienů z mitochondrie. Tyto se účastní na kaspázách nezávislé jaderné degradace DNA, zejména pak protein AIF (AIF, označovaný také jako PDCD8 nebo AIFM1) a endonukleáza G. Další zajímavý protein, který však není v mitochondriích, ale předpokládá se jeho podobná funkce jakou má AIF, neboť s tímto proteinem sdílí sekvenční homologii. Tímto proteinem je AIF-homologous mitochondrion-associated inducer of death (AMID, též PRG3 nebo AIFM2). Studovali jsem lokalizaci a interakce těchto proteinů v živých lidských buňkách. Naše výsledky značně přispěly k pochopení lokalizace, interakcí a funkce proteinů AMID, AIF a endonukleázy G v lidských buňkách.
Links
| GP204/05/P090, research and development project |
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| LC535, research and development project |
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| MSM0021622419, plan (intention) |
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