Bioinformatic and image analyses of the cellular localization of the apoptotic proteins endonuclease G, AIF, and AMID during apoptosis in human cells.
We studied the cellular localization of the apoptotic proteins endonuclease G, AIF, and AMID in silico using three prediction tools and in living cells using both single-cell colocalization image analysis and nuclear translocation analysis. We confirmed the mitochondrial localization of endonuclease G and AIF by prediction analysis and by single-cell colocalization image analysis. We found the AMID protein to be cytoplasmic, most probably incorporated into the cytoplasmic side of the membranes of various organelles. The highest concentration of AMID was observed associated with the Golgi. Colocalization of AMID with lysosomes was also indirectly confirmed by analysis of AMID-rich vesicle velocity using manual tracking analysis. Bioinformatic analysis also detected nuclear localization signals in endonuclease G and AIF, but not in AMID. A novel analysis of time-lapse fluorescence image data during staurosporine-induced apoptosis revealed nuclear translocation only for endonuclease G and AIF.
Varecha
Miroslav
M
Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Botanická 68a, Brno, 602 00, Czech Republic. mvara@fi.muni.cz
Amrichová
Jana
J
Zimmermann
Michal
M
Ulman
Vladimír
V
Lukásová
Emilie
E
Kozubek
Michal
M
D017209:Apoptosis / Q000502:physiology*; D051033:Apoptosis Inducing Factor / Q000302:isolation & purification / Q000378:metabolism*; D051017:Apoptosis Regulatory Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D019295:Computational Biology; D004706:Endodeoxyribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
C494829:AIFM1 protein, human; C462504:ferroptosis suppressor protein 1, human; D051033:Apoptosis Inducing Factor; D051017:Apoptosis Regulatory Proteins; D024101:Mitochondrial Proteins; D011993:Recombinant Fusion Proteins; D004706:Endodeoxyribonucleases; C052391:endonuclease G