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.
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Research Article|
March 15 2011
Light-induced blockage of cell division with a chromatin-targeted phototoxic fluorescent protein
Ekaterina O. Serebrovskaya;
Ekaterina O. Serebrovskaya
1
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
†Nizhny Novgorod State Medical Academy, 10/1 Minin and Pozharsky Square, 603005 Nizhny Novgorod, Russia
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Tatiana V. Gorodnicheva;
Tatiana V. Gorodnicheva
2
‡Evrogen JSC, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
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Galina V. Ermakova;
Galina V. Ermakova
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
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Elena A. Solovieva;
Elena A. Solovieva
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
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George V. Sharonov;
George V. Sharonov
§Faculty of Medicine, Lomonosov Moscow State University, 119192 Moscow, Russia
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Elena V. Zagaynova;
Elena V. Zagaynova
†Nizhny Novgorod State Medical Academy, 10/1 Minin and Pozharsky Square, 603005 Nizhny Novgorod, Russia
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Dmitriy M. Chudakov;
Dmitriy M. Chudakov
3
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
†Nizhny Novgorod State Medical Academy, 10/1 Minin and Pozharsky Square, 603005 Nizhny Novgorod, Russia
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Sergey Lukyanov;
Sergey Lukyanov
3
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
†Nizhny Novgorod State Medical Academy, 10/1 Minin and Pozharsky Square, 603005 Nizhny Novgorod, Russia
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Andrey G. Zaraisky;
Andrey G. Zaraisky
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
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Konstantin A. Lukyanov
*Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997 Moscow, Russia
†Nizhny Novgorod State Medical Academy, 10/1 Minin and Pozharsky Square, 603005 Nizhny Novgorod, Russia
4To whom correspondence should be addressed (email kluk@ibch.ru).
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Publisher: Portland Press Ltd
Received:
August 04 2010
Revision Received:
January 06 2011
Accepted:
January 10 2011
Accepted Manuscript online:
January 10 2011
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2011 Biochemical Society
2011
Biochem J (2011) 435 (1): 65–71.
Article history
Received:
August 04 2010
Revision Received:
January 06 2011
Accepted:
January 10 2011
Accepted Manuscript online:
January 10 2011
Citation
Ekaterina O. Serebrovskaya, Tatiana V. Gorodnicheva, Galina V. Ermakova, Elena A. Solovieva, George V. Sharonov, Elena V. Zagaynova, Dmitriy M. Chudakov, Sergey Lukyanov, Andrey G. Zaraisky, Konstantin A. Lukyanov; Light-induced blockage of cell division with a chromatin-targeted phototoxic fluorescent protein. Biochem J 1 April 2011; 435 (1): 65–71. doi: https://doi.org/10.1042/BJ20101217
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