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Dynamic changes in subcellular localization of cattle XLF during cell cycle, and focus formation of cattle XLF at DNA damage sites immediately after irradiation.

Koike M, Yutoku Y, Koike A - J. Vet. Med. Sci. (2015)

Bottom Line: Moreover, nuclear localization and accumulation of cattle XLF at DSB sites are dependent on 12 amino acids (288-299) of the C-terminal region of XLF (XLF CTR).Furthermore, basic amino acids on the XLF CTR are highly conserved among domestic animals including cattle, goat and horses, suggesting that the CTR is essential for the function of XLF in domestic animals.These findings might be useful to develop the molecular-targeting therapeutic drug taking XLF as a target molecule for human and domestic animals.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555, Japan.

ABSTRACT
Clinically, many chemotherapeutics and ionizing radiation (IR) have been applied for the treatment of various types of human and animal malignancies. These treatments kill tumor cells by causing DNA double-strand breaks (DSBs). Core factors of classical nonhomologous DNA-end joining (C-NHEJ) play a vital role in DSB repair. Thus, it is indispensable to clarify the mechanisms of C-NHEJ in order to develop next-generation chemotherapeutics for cancer. The XRCC4-like factor (XLF; also called Cernunnos or NHEJ1) is the lastly identified core NHEJ factor. The localization of core NHEJ factors might play a critical role in regulating NHEJ activity. The localization and function of XLF have not been elucidated in animal species other than mice and humans. Domestic cattle (Bos taurus) are the most common and vital domestic animals in many countries. Here, we show that the localization of cattle XLF changes dynamically during the cell cycle. Furthermore, EYFP-cattle XLF accumulates quickly at microirradiated sites and colocalizes with the DSB marker γH2AX. Moreover, nuclear localization and accumulation of cattle XLF at DSB sites are dependent on 12 amino acids (288-299) of the C-terminal region of XLF (XLF CTR). Furthermore, basic amino acids on the XLF CTR are highly conserved among domestic animals including cattle, goat and horses, suggesting that the CTR is essential for the function of XLF in domestic animals. These findings might be useful to develop the molecular-targeting therapeutic drug taking XLF as a target molecule for human and domestic animals.

No MeSH data available.


Related in: MedlinePlus

EYFP-cattle XLF accumulated quickly at DSBs induced by laser microirradiation. (A)The localization and accumulation of EYFP-cattle XLF at DSBs induced by 405 nm laserirradiation were examined. (B) Imaging of living EYFP-cattle XLF-transfected MDBKcells before (upper panel) and at 1 min after (lower panel) microirradiation. Leftpanel, EYFP-cattle XLF; right panel, differential interference contrast images (DIC).Arrowheads indicate the microirradiated sites. (C) Immunostaining of microirradiatedEYFP-cattle XLF-transfected cells with anti-γH2AX antibody. The cells were fixed andstained with the anti- γH2AX antibody at 5 min postirradiation. Left panel,EYFP-cattle XLF; center panel, γH2AX image; right panel, merged image. (D)Time-dependent EYFP-cattle XLF accumulation in living cells (5-120 sec) afterirradiation. Upper panel, EYFP-cattle XLF; lower panel, differential interferencecontrast images (DIC).
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fig_003: EYFP-cattle XLF accumulated quickly at DSBs induced by laser microirradiation. (A)The localization and accumulation of EYFP-cattle XLF at DSBs induced by 405 nm laserirradiation were examined. (B) Imaging of living EYFP-cattle XLF-transfected MDBKcells before (upper panel) and at 1 min after (lower panel) microirradiation. Leftpanel, EYFP-cattle XLF; right panel, differential interference contrast images (DIC).Arrowheads indicate the microirradiated sites. (C) Immunostaining of microirradiatedEYFP-cattle XLF-transfected cells with anti-γH2AX antibody. The cells were fixed andstained with the anti- γH2AX antibody at 5 min postirradiation. Left panel,EYFP-cattle XLF; center panel, γH2AX image; right panel, merged image. (D)Time-dependent EYFP-cattle XLF accumulation in living cells (5-120 sec) afterirradiation. Upper panel, EYFP-cattle XLF; lower panel, differential interferencecontrast images (DIC).

Mentions: EYFP-cattle XLF accumulates quickly at DSBs induced by lasermicroirradiation: We examined whether EYFP-cattle XLF accumulates quickly at the405 nm laser-induced DSB sites (Fig. 3AFig. 3.


Dynamic changes in subcellular localization of cattle XLF during cell cycle, and focus formation of cattle XLF at DNA damage sites immediately after irradiation.

Koike M, Yutoku Y, Koike A - J. Vet. Med. Sci. (2015)

EYFP-cattle XLF accumulated quickly at DSBs induced by laser microirradiation. (A)The localization and accumulation of EYFP-cattle XLF at DSBs induced by 405 nm laserirradiation were examined. (B) Imaging of living EYFP-cattle XLF-transfected MDBKcells before (upper panel) and at 1 min after (lower panel) microirradiation. Leftpanel, EYFP-cattle XLF; right panel, differential interference contrast images (DIC).Arrowheads indicate the microirradiated sites. (C) Immunostaining of microirradiatedEYFP-cattle XLF-transfected cells with anti-γH2AX antibody. The cells were fixed andstained with the anti- γH2AX antibody at 5 min postirradiation. Left panel,EYFP-cattle XLF; center panel, γH2AX image; right panel, merged image. (D)Time-dependent EYFP-cattle XLF accumulation in living cells (5-120 sec) afterirradiation. Upper panel, EYFP-cattle XLF; lower panel, differential interferencecontrast images (DIC).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591152&req=5

fig_003: EYFP-cattle XLF accumulated quickly at DSBs induced by laser microirradiation. (A)The localization and accumulation of EYFP-cattle XLF at DSBs induced by 405 nm laserirradiation were examined. (B) Imaging of living EYFP-cattle XLF-transfected MDBKcells before (upper panel) and at 1 min after (lower panel) microirradiation. Leftpanel, EYFP-cattle XLF; right panel, differential interference contrast images (DIC).Arrowheads indicate the microirradiated sites. (C) Immunostaining of microirradiatedEYFP-cattle XLF-transfected cells with anti-γH2AX antibody. The cells were fixed andstained with the anti- γH2AX antibody at 5 min postirradiation. Left panel,EYFP-cattle XLF; center panel, γH2AX image; right panel, merged image. (D)Time-dependent EYFP-cattle XLF accumulation in living cells (5-120 sec) afterirradiation. Upper panel, EYFP-cattle XLF; lower panel, differential interferencecontrast images (DIC).
Mentions: EYFP-cattle XLF accumulates quickly at DSBs induced by lasermicroirradiation: We examined whether EYFP-cattle XLF accumulates quickly at the405 nm laser-induced DSB sites (Fig. 3AFig. 3.

Bottom Line: Moreover, nuclear localization and accumulation of cattle XLF at DSB sites are dependent on 12 amino acids (288-299) of the C-terminal region of XLF (XLF CTR).Furthermore, basic amino acids on the XLF CTR are highly conserved among domestic animals including cattle, goat and horses, suggesting that the CTR is essential for the function of XLF in domestic animals.These findings might be useful to develop the molecular-targeting therapeutic drug taking XLF as a target molecule for human and domestic animals.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555, Japan.

ABSTRACT
Clinically, many chemotherapeutics and ionizing radiation (IR) have been applied for the treatment of various types of human and animal malignancies. These treatments kill tumor cells by causing DNA double-strand breaks (DSBs). Core factors of classical nonhomologous DNA-end joining (C-NHEJ) play a vital role in DSB repair. Thus, it is indispensable to clarify the mechanisms of C-NHEJ in order to develop next-generation chemotherapeutics for cancer. The XRCC4-like factor (XLF; also called Cernunnos or NHEJ1) is the lastly identified core NHEJ factor. The localization of core NHEJ factors might play a critical role in regulating NHEJ activity. The localization and function of XLF have not been elucidated in animal species other than mice and humans. Domestic cattle (Bos taurus) are the most common and vital domestic animals in many countries. Here, we show that the localization of cattle XLF changes dynamically during the cell cycle. Furthermore, EYFP-cattle XLF accumulates quickly at microirradiated sites and colocalizes with the DSB marker γH2AX. Moreover, nuclear localization and accumulation of cattle XLF at DSB sites are dependent on 12 amino acids (288-299) of the C-terminal region of XLF (XLF CTR). Furthermore, basic amino acids on the XLF CTR are highly conserved among domestic animals including cattle, goat and horses, suggesting that the CTR is essential for the function of XLF in domestic animals. These findings might be useful to develop the molecular-targeting therapeutic drug taking XLF as a target molecule for human and domestic animals.

No MeSH data available.


Related in: MedlinePlus