title: Light-induced blockage of cell division with a chromatin-targeted phototoxic fluorescent protein.
abstract: Proteins of the GFP (green fluorescent protein) family are widely used as passive reporters for live cell imaging. In the present study we used H2B (histone H2B)-tKR (tandem KillerRed) as an active tool to affect cell division with light. We demonstrated that H2B-tKR-expressing cells behave normally in the dark, but transiently cease proliferation following green-light illumination. Complete light-induced blockage of cell division for approx. 24 h was observed in cultured mammalian cells that were either transiently or stably transfected with H2B-tKR. Illuminated cells then returned to normal division rate. XRCC1 (X-ray cross complementing factor 1) showed immediate redistribution in the illuminated nuclei of H2B-tKR-expressing cells, indicating massive light-induced damage of genomic DNA. Notably, nondisjunction of chromosomes was observed for cells that were illuminated during metaphase. In transgenic Xenopus embryos expressing H2B-tKR under the control of tissue-specific promoters, we observed clear retardation of the development of these tissues in green-light-illuminated tadpoles. We believe that H2B-tKR represents a novel optogenetic tool, which can be used to study mitosis and meiosis progression per se, as well as to investigate the roles of specific cell populations in development, regeneration and carcinogenesis in vivo.
authors.affiliation: Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia.
authors.forename: Ekaterina O
authors.initials: EO
authors.lastname: Serebrovskaya
authors.forename: Tatiana V
authors.initials: TV
authors.lastname: Gorodnicheva
authors.forename: Galina V
authors.initials: GV
authors.lastname: Ermakova
authors.forename: Elena A
authors.initials: EA
authors.lastname: Solovieva
authors.forename: George V
authors.initials: GV
authors.lastname: Sharonov
authors.forename: Elena V
authors.initials: EV
authors.lastname: Zagaynova
authors.forename: Dmitriy M
authors.initials: DM
authors.lastname: Chudakov
authors.forename: Sergey
authors.initials: S
authors.lastname: Lukyanov
authors.forename: Andrey G
authors.initials: AG
authors.lastname: Zaraisky
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Lukyanov
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology / Q000378:metabolism; D002455:Cell Division / Q000528:radiation effects*; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism* / Q000528:radiation effects; D004249:DNA Damage / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008027:Light; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D015335:Molecular Probes / Q000235:genetics / Q000378:metabolism*; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D021381:Protein Transport / Q000528:radiation effects; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D000076105:X-ray Repair Cross Complementing Protein 1; D014982:Xenopus laevis
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D006657:Histones; D008164:Luminescent Proteins; D015335:Molecular Probes; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D000076105:X-ray Repair Cross Complementing Protein 1; C000619371:XRCC1 protein, human; C558184:histone H2B-tandem killer red protein, human; C508771:killer red protein, Anthomedusae; D049452:Green Fluorescent Proteins