Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C
Deinococcus radiodurans known for its extraordinary resistance to ionizing radiation contains bacterial phytochrome (BphP), a member of the family of red/far-red light-sensing proteins. In this study, we constructed a bphP mutant strain (ΔbphP) to investigate the role of D. radiodurans BphP (DrBphP)...
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2021-07-01
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doaj-44b020be77af4fe5929251fcac9554fa2021-07-30T06:15:26ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-07-011210.3389/fmicb.2021.659233659233Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin CJong-Hyun Jung0Jong-Hyun Jung1Soyoung Jeong2Soyoung Jeong3Seonghun Im4Min-Kyu Kim5Ho Seong Seo6Ho Seong Seo7Sangyong Lim8Sangyong Lim9Radiation Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaDepartment of Radiation Science and Technology, University of Science and Technology, Daejeon, South KoreaRadiation Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaDepartment of Food and Animal Biotechnology, Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, South KoreaRadiation Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaRadiation Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaRadiation Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaDepartment of Radiation Science and Technology, University of Science and Technology, Daejeon, South KoreaRadiation Research Division, Korea Atomic Energy Research Institute, Jeongeup, South KoreaDepartment of Radiation Science and Technology, University of Science and Technology, Daejeon, South KoreaDeinococcus radiodurans known for its extraordinary resistance to ionizing radiation contains bacterial phytochrome (BphP), a member of the family of red/far-red light-sensing proteins. In this study, we constructed a bphP mutant strain (ΔbphP) to investigate the role of D. radiodurans BphP (DrBphP) in the DNA damage response. When cells were incubated under light and dark conditions following exposure to DNA damaging agents, such as γ- and UV-radiation and mitomycin C (MMC), no significant difference in cell survival was observed between the wild-type D. radiodurans strain (WT) and ΔbphP. However, when continuously exposed to MMC under light conditions, the WT strain notably exhibited increased survival compared to cells grown in the dark. The increased survival was not observed in the ΔbphP strain. These results are indicative of the protective role of light-activated DrBphP in the presence of MMC. Site-directed mutagenesis revealed that the conserved amino acids Cys-24 and His-532 involved in chromophore binding and signal transduction, respectively, were essential for the protective function of DrBphP. Inactivation of the cognate response regulator (RR; DrBphR) of DrBphP increased MMC resistance in the dark. In trans complementation of the bphP bphR double mutant strain (ΔbphPR) with DrBphR decreased MMC resistance. Considering that DrBphP acts as a light-activated phosphatase that dephosphorylates DrBphR, it appears that phosphorylated DrBphR exerts a negative effect on cell survival in the presence of MMC. DrBphP overexpression resulted in an increase in MMC resistance of ΔbphPR, implying that other RRs might be involved in the DrBphP-mediated signaling pathway. A mutant lacking the dr_0781 gene (Δdr_0781) demonstrated the same MMC phenotype as ΔbphR. Survival was further increased in the bphR dr_0781 double mutant strain compared to each single mutant ΔbphR or Δdr_0781, suggesting that DR_0781 is also involved in the DrBphP-dependent MMC sensitivity. This study uncovered a previously unknown phenomenon of red/far-red light-dependent DNA damage survival mediated by BphP by identifying the conditions under which DrBphP exhibits a fitness advantage.https://www.frontiersin.org/articles/10.3389/fmicb.2021.659233/fullDeinococcus radioduransbacteriophytochromeBphPBphRlight illuminationmitomycin C resistance |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jong-Hyun Jung Jong-Hyun Jung Soyoung Jeong Soyoung Jeong Seonghun Im Min-Kyu Kim Ho Seong Seo Ho Seong Seo Sangyong Lim Sangyong Lim |
spellingShingle |
Jong-Hyun Jung Jong-Hyun Jung Soyoung Jeong Soyoung Jeong Seonghun Im Min-Kyu Kim Ho Seong Seo Ho Seong Seo Sangyong Lim Sangyong Lim Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C Frontiers in Microbiology Deinococcus radiodurans bacteriophytochrome BphP BphR light illumination mitomycin C resistance |
author_facet |
Jong-Hyun Jung Jong-Hyun Jung Soyoung Jeong Soyoung Jeong Seonghun Im Min-Kyu Kim Ho Seong Seo Ho Seong Seo Sangyong Lim Sangyong Lim |
author_sort |
Jong-Hyun Jung |
title |
Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C |
title_short |
Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C |
title_full |
Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C |
title_fullStr |
Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C |
title_full_unstemmed |
Lack of the Bacterial Phytochrome Protein Decreases Deinococcus radiodurans Resistance to Mitomycin C |
title_sort |
lack of the bacterial phytochrome protein decreases deinococcus radiodurans resistance to mitomycin c |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Microbiology |
issn |
1664-302X |
publishDate |
2021-07-01 |
description |
Deinococcus radiodurans known for its extraordinary resistance to ionizing radiation contains bacterial phytochrome (BphP), a member of the family of red/far-red light-sensing proteins. In this study, we constructed a bphP mutant strain (ΔbphP) to investigate the role of D. radiodurans BphP (DrBphP) in the DNA damage response. When cells were incubated under light and dark conditions following exposure to DNA damaging agents, such as γ- and UV-radiation and mitomycin C (MMC), no significant difference in cell survival was observed between the wild-type D. radiodurans strain (WT) and ΔbphP. However, when continuously exposed to MMC under light conditions, the WT strain notably exhibited increased survival compared to cells grown in the dark. The increased survival was not observed in the ΔbphP strain. These results are indicative of the protective role of light-activated DrBphP in the presence of MMC. Site-directed mutagenesis revealed that the conserved amino acids Cys-24 and His-532 involved in chromophore binding and signal transduction, respectively, were essential for the protective function of DrBphP. Inactivation of the cognate response regulator (RR; DrBphR) of DrBphP increased MMC resistance in the dark. In trans complementation of the bphP bphR double mutant strain (ΔbphPR) with DrBphR decreased MMC resistance. Considering that DrBphP acts as a light-activated phosphatase that dephosphorylates DrBphR, it appears that phosphorylated DrBphR exerts a negative effect on cell survival in the presence of MMC. DrBphP overexpression resulted in an increase in MMC resistance of ΔbphPR, implying that other RRs might be involved in the DrBphP-mediated signaling pathway. A mutant lacking the dr_0781 gene (Δdr_0781) demonstrated the same MMC phenotype as ΔbphR. Survival was further increased in the bphR dr_0781 double mutant strain compared to each single mutant ΔbphR or Δdr_0781, suggesting that DR_0781 is also involved in the DrBphP-dependent MMC sensitivity. This study uncovered a previously unknown phenomenon of red/far-red light-dependent DNA damage survival mediated by BphP by identifying the conditions under which DrBphP exhibits a fitness advantage. |
topic |
Deinococcus radiodurans bacteriophytochrome BphP BphR light illumination mitomycin C resistance |
url |
https://www.frontiersin.org/articles/10.3389/fmicb.2021.659233/full |
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