RecQ is a highly conserved helicase necessary for maintaining genome stability in all organisms. Genome comparison showed that a homologue of RecQ in Deinococcus radiodurans designated as DR1289 is a member of RecQ fa...RecQ is a highly conserved helicase necessary for maintaining genome stability in all organisms. Genome comparison showed that a homologue of RecQ in Deinococcus radiodurans designated as DR1289 is a member of RecQ family with unusual domain arrangement: a helicase domain, an RecQ C-terminal domain, and surprisingly three HRDC domain repeats, whose func-tion, however, remains obscure currently. Using an insertion deletion, we discovered that the DRRecQ mutation causes an increase in gamma radiation, hydroxyurea and mitomycine C and UV sensitivity. Using the shuttle plasmid pRADK, we complemented various domains of the D. radiodurans RecQ (DRRecQ) to the mutant in vivo. Results suggested that both the helicase and helicase-and-RNase-D-C-terminal (HRDC) domains are essential for complementing several phenotypes. The complementation and biochemical function of DRRecQ variants with different domains truncated in vitro suggested that both the helicase and three HRDC domains are necessary for RecQ functions in D. radiodurans, while three HRDC domains have a synergistic effect on the whole function. Our finding leads to the hypothesis that the RecF recombination pathway is likely a primary path of double strand break repair in this well-known radioresistant organism.展开更多
To explore the survival and dose response of organism for different radiation sources is of great importance in the research of radiobiology. In this study, the survival-dose response of Deinococcus radiodurans (E.col...To explore the survival and dose response of organism for different radiation sources is of great importance in the research of radiobiology. In this study, the survival-dose response of Deinococcus radiodurans (E.coli, as the control) for ultra-violet (UV), γ-rays radiation and ion beam exposure was investigated. The shoulder type of survival curves were found for both UV and γ-ray ionizing radiation, but the saddle type of survival curves were shown for H+、 N+( 20keV and 30keV) and Ar+ beam exposure. This dose effect of the survival initially decreased with the increase in dose and then increased in the high dose range and finally decreased again in the higher dose range. Our experimental results suggest that D. radiodurans, which is considerably radio-resistant to UV and x-ray and γ-ray ionizing radiation, do not resist ion beam exposure.展开更多
Though bacteria of the radiation-resistant Deinococcus radiodurans have a high resistance to the lethal and mutagenic effects of many DNA-damaging agents, the mechanisms involved in the response of these bacteria to o...Though bacteria of the radiation-resistant Deinococcus radiodurans have a high resistance to the lethal and mutagenic effects of many DNA-damaging agents, the mechanisms involved in the response of these bacteria to oxidative stress are poorly understood. In this report, the superoxide dismutase (SOD) and catalase (CAT) activities produced by these bacteria were measured, and the change of SOD and CAT activities by 20 keV N+ beam exposure was examined. Their activities were increased by N+ beam exposure from 8x 1014 ions/cm2 to 6x1015 ions/cm2.The treatment of H2O2 and [ CHC13 +CH3 CH2OH ] and the measurement of absorption spectrum showed that the increase in SOD activity was resulted from inducible activities of MnSOD in D. radiodurans AS1.633 by N+ beam exposure . These results suggested that this bacteria possess inducible defense mechanisms against the deleterious effects of oxidization.展开更多
Objectives:This study used Deinococcus radiodurans,which is extremely resistant to oxidative damage,genotoxic chemicals,high levels of ionising and ultraviolet radiation and drying,and its Vitreoscilla haemoglobin(vgb...Objectives:This study used Deinococcus radiodurans,which is extremely resistant to oxidative damage,genotoxic chemicals,high levels of ionising and ultraviolet radiation and drying,and its Vitreoscilla haemoglobin(vgb)gene-cloned recombinant with the vgb−recombinant strain as a control.In addition to the conditions wherein bacteria have an optimum Cr(Ⅲ)biosorption capacity,the contribution of the vgb gene to the biosorption ability of the bacteria has been investigated by providing the organism with a more oxygenic environment.Methods:Bacteria were produced and metal stock solution was prepared.To determine the Cr(Ⅲ)removal capacities of wild and recombinant D.radiodurans strains,the residual metal concentration in aqueous media at the beginning and after biosorption was determined in Atomic Absorption Spectrophotometer.Some optimal conditions were created for the biosorption conditions to occur.Conclusions:The optimisation tests showed that Cr(Ⅲ)reached the highest biosorption capacity within 15 minutes at a metal concentration of 2,000 ppm,30°C,pH 5.0 and 150 rpm stirring speed in all the three bacteria.The vgb gene had no significant contribution to the biosorption capacity.展开更多
Deinococcus radiodurans (D. radiodurans) is distinguished by the most radioresistant organism ever known, and can tolerate extreme environments such as ionizing radiation, ultraviolet radiation, oxidation, and desicca...Deinococcus radiodurans (D. radiodurans) is distinguished by the most radioresistant organism ever known, and can tolerate extreme environments such as ionizing radiation, ultraviolet radiation, oxidation, and desiccation. D. radiodurans is an important model for studying DNA damage/repair and redox regulation upon high dose ionizing radiation. How D. radiodurans response and repair ROS-induced oxidative damage remains a subject of ongoing investigation. This review provides an overview of the radioresistance characteristics of D. radiodurans. Among them, the DNA damage repair pathway and high-efficiency antioxidant defense system are summarized in detail. Furthermore, a novel model that protects the cell against the ionizing radiation is proposed. This review also discusses the potential application , future challenges and directions in advancing towards D. radiodurans studies.展开更多
PprI, a DNA damage response factor from the extraordinary radioresistant bacterium Deinococcus radi- odurans, plays a central regulatory role in multiple DNA damage repair. In this study, a fusion DNA fragment carry- ...PprI, a DNA damage response factor from the extraordinary radioresistant bacterium Deinococcus radi- odurans, plays a central regulatory role in multiple DNA damage repair. In this study, a fusion DNA fragment carry- ing kanamycin resistance gene with the D. radiodurans groEL promoter was cloned by PCR amplification and reversely inserted into the pprI locus in the genome of the wild-type strain R1. The resulting pprI-deficient strain, designated YR1, was very sensitive to ionizing radiation. Meanwhile, the re- combinant DNA fragment was cloned into the shuttle vector pRADZ3, and resulted in plasmid pRADK with kanamycin resistance in D. radiodurans. The fragments containing com- plete pprI gene and 3′-terminal deletion pprI△ were cloned into plasmid pRADK. The resulted plasmids designated pRADKpprI and pRADKpprI△ were then transformed to YR1. Results show that YR1 carrying pRADKpprI was able to fully restore the extreme radioresistance to the same level as the wild-type D. raiodurans R1, whereas YR1 pRADKpprI△ failed to do so. Construction of DNA repair switch PprI function-deficient and function-complementary mutants in D. radiodurans is not only useful to elucidating the relationship between domains and functions of PprI pro- tein, but also opens the door to the further studies of the bio- logical functions of PprI protein in vivo.展开更多
The conclusion based on transmission electron microscopy, "the tightly packed ring-like nucleoid of the Deinococcus radiodurans R1 is a key to radioresistance", has instigated lots of debates. In this study,...The conclusion based on transmission electron microscopy, "the tightly packed ring-like nucleoid of the Deinococcus radiodurans R1 is a key to radioresistance", has instigated lots of debates. In this study, according to the previous research of PprI’s crucial role in radioresistance of D. radiodurans, we have attempted to examine and compare the nucleoid morphology differences among wild-type D. ra-diodurans R1 strain, pprI function-deficient mutant (YR1), and pprI function-complementary strains (YR1001, YR1002, and YR1004) before and after exposure to ionizing irradiation. Fluorescence mi-croscopy images indicate: (1) the majority of nucleoid structures in radioresistant strain R1 cells ex-hibit the tightly packed ring-like morphology, while the pprI function-deficient mutant YR1 cells carrying predominate ring-like structure represent high sensitivity to irradiation; (2) as an extreme radioresistant strain similar to wild-type R1, pprI completely function-complementary strain YR1001 almost displays the loose and irregular nucleoid morphologies. On the other hand, another radioresistant pprI partly function-complementary strain YR1002’s nucleiods exhibit about 60% ring-like structure; (3) a PprI C-terminal deletion strain YR1004 consisting of approximately 60% of ring-like nucleoid is very sensi-tive to radiation. Therefore, our present experiments do not support the conclusion that the ring-like nucleoid of D. radiodurans does play a key role in radioresistance.展开更多
The β-subunit of RNA polymerase, which is involved in rifampin binding, is highly conserved among prokaryotes, and Rile mutants detected in many bacteria are the result of amino acid changes. Spontaneous rifampin res...The β-subunit of RNA polymerase, which is involved in rifampin binding, is highly conserved among prokaryotes, and Rile mutants detected in many bacteria are the result of amino acid changes. Spontaneous rifampin resistance mutations resulting in amino acid replacement （L420R） and deletion （1258-66 9 bp deletion） have been previously isolated in the rpoB gene of Deinococcus radiodurans. In this study, a β-subunit mutation in D. radiodurans resulted in a unique effect on growth rate. We used DNA microarrays and biochemical assays to investigate how the Rif mutation in the β-subunit led to changes in growth rate via altered regulation of multiple genes. The expression of genes with predicted functions in metabolism, cellular processes and signaling, and information storage and processing were significantly altered in the 9 bp-deletion rpoB mutant. The consensus promoter sequence of up-regulated genes in the 9 bp-deletion rpoB mutant was identified as an AT-rich sequence. Greater levels of reactive oxygen species accumulated in the L420R and 9 bp-deletion rpoB mutants compared with wild type. These results provide insight into the molecular mechanism of how the β-subunit Rif mutation alters the regulation of multiple genes.展开更多
基金Project supported by the National Basic Research Program (973) of China (No. 2004CB19604), Distinguished Young Scientist of China (No. 30425038), and the National Natural Science Foundation of China (No. 30330020)
文摘RecQ is a highly conserved helicase necessary for maintaining genome stability in all organisms. Genome comparison showed that a homologue of RecQ in Deinococcus radiodurans designated as DR1289 is a member of RecQ family with unusual domain arrangement: a helicase domain, an RecQ C-terminal domain, and surprisingly three HRDC domain repeats, whose func-tion, however, remains obscure currently. Using an insertion deletion, we discovered that the DRRecQ mutation causes an increase in gamma radiation, hydroxyurea and mitomycine C and UV sensitivity. Using the shuttle plasmid pRADK, we complemented various domains of the D. radiodurans RecQ (DRRecQ) to the mutant in vivo. Results suggested that both the helicase and helicase-and-RNase-D-C-terminal (HRDC) domains are essential for complementing several phenotypes. The complementation and biochemical function of DRRecQ variants with different domains truncated in vitro suggested that both the helicase and three HRDC domains are necessary for RecQ functions in D. radiodurans, while three HRDC domains have a synergistic effect on the whole function. Our finding leads to the hypothesis that the RecF recombination pathway is likely a primary path of double strand break repair in this well-known radioresistant organism.
基金the National Natural Science Foundation of China! No.196O5005)
文摘To explore the survival and dose response of organism for different radiation sources is of great importance in the research of radiobiology. In this study, the survival-dose response of Deinococcus radiodurans (E.coli, as the control) for ultra-violet (UV), γ-rays radiation and ion beam exposure was investigated. The shoulder type of survival curves were found for both UV and γ-ray ionizing radiation, but the saddle type of survival curves were shown for H+、 N+( 20keV and 30keV) and Ar+ beam exposure. This dose effect of the survival initially decreased with the increase in dose and then increased in the high dose range and finally decreased again in the higher dose range. Our experimental results suggest that D. radiodurans, which is considerably radio-resistant to UV and x-ray and γ-ray ionizing radiation, do not resist ion beam exposure.
基金This work was supported by the National Nature Science Foundation of China! No.19605005
文摘Though bacteria of the radiation-resistant Deinococcus radiodurans have a high resistance to the lethal and mutagenic effects of many DNA-damaging agents, the mechanisms involved in the response of these bacteria to oxidative stress are poorly understood. In this report, the superoxide dismutase (SOD) and catalase (CAT) activities produced by these bacteria were measured, and the change of SOD and CAT activities by 20 keV N+ beam exposure was examined. Their activities were increased by N+ beam exposure from 8x 1014 ions/cm2 to 6x1015 ions/cm2.The treatment of H2O2 and [ CHC13 +CH3 CH2OH ] and the measurement of absorption spectrum showed that the increase in SOD activity was resulted from inducible activities of MnSOD in D. radiodurans AS1.633 by N+ beam exposure . These results suggested that this bacteria possess inducible defense mechanisms against the deleterious effects of oxidization.
基金This study was supported by Inonu University Scientific Research Projects Coordination Unit(project number:2012-192).
文摘Objectives:This study used Deinococcus radiodurans,which is extremely resistant to oxidative damage,genotoxic chemicals,high levels of ionising and ultraviolet radiation and drying,and its Vitreoscilla haemoglobin(vgb)gene-cloned recombinant with the vgb−recombinant strain as a control.In addition to the conditions wherein bacteria have an optimum Cr(Ⅲ)biosorption capacity,the contribution of the vgb gene to the biosorption ability of the bacteria has been investigated by providing the organism with a more oxygenic environment.Methods:Bacteria were produced and metal stock solution was prepared.To determine the Cr(Ⅲ)removal capacities of wild and recombinant D.radiodurans strains,the residual metal concentration in aqueous media at the beginning and after biosorption was determined in Atomic Absorption Spectrophotometer.Some optimal conditions were created for the biosorption conditions to occur.Conclusions:The optimisation tests showed that Cr(Ⅲ)reached the highest biosorption capacity within 15 minutes at a metal concentration of 2,000 ppm,30°C,pH 5.0 and 150 rpm stirring speed in all the three bacteria.The vgb gene had no significant contribution to the biosorption capacity.
基金supported by the National Undergraduate Training Program for Innovation and Entrepreneurship of China(Project number:202010007748).
文摘Deinococcus radiodurans (D. radiodurans) is distinguished by the most radioresistant organism ever known, and can tolerate extreme environments such as ionizing radiation, ultraviolet radiation, oxidation, and desiccation. D. radiodurans is an important model for studying DNA damage/repair and redox regulation upon high dose ionizing radiation. How D. radiodurans response and repair ROS-induced oxidative damage remains a subject of ongoing investigation. This review provides an overview of the radioresistance characteristics of D. radiodurans. Among them, the DNA damage repair pathway and high-efficiency antioxidant defense system are summarized in detail. Furthermore, a novel model that protects the cell against the ionizing radiation is proposed. This review also discusses the potential application , future challenges and directions in advancing towards D. radiodurans studies.
基金This work was supported by the National Basic Research Program of China(Grant No.2004CB 19604)the Distin-guished Young Scientists of China(Grant No.30425038)the Na-tional Natural Science Foundation of China(Grant No.30330020).
文摘PprI, a DNA damage response factor from the extraordinary radioresistant bacterium Deinococcus radi- odurans, plays a central regulatory role in multiple DNA damage repair. In this study, a fusion DNA fragment carry- ing kanamycin resistance gene with the D. radiodurans groEL promoter was cloned by PCR amplification and reversely inserted into the pprI locus in the genome of the wild-type strain R1. The resulting pprI-deficient strain, designated YR1, was very sensitive to ionizing radiation. Meanwhile, the re- combinant DNA fragment was cloned into the shuttle vector pRADZ3, and resulted in plasmid pRADK with kanamycin resistance in D. radiodurans. The fragments containing com- plete pprI gene and 3′-terminal deletion pprI△ were cloned into plasmid pRADK. The resulted plasmids designated pRADKpprI and pRADKpprI△ were then transformed to YR1. Results show that YR1 carrying pRADKpprI was able to fully restore the extreme radioresistance to the same level as the wild-type D. raiodurans R1, whereas YR1 pRADKpprI△ failed to do so. Construction of DNA repair switch PprI function-deficient and function-complementary mutants in D. radiodurans is not only useful to elucidating the relationship between domains and functions of PprI pro- tein, but also opens the door to the further studies of the bio- logical functions of PprI protein in vivo.
基金Supported by the National Basic Research Program of China (Grant No. 2004CB19604)the National Natural Science Foundation of China (Grant No. 30330020)the National Science fund for Distinguished Young Scholars (Grant No. 30425038)
文摘The conclusion based on transmission electron microscopy, "the tightly packed ring-like nucleoid of the Deinococcus radiodurans R1 is a key to radioresistance", has instigated lots of debates. In this study, according to the previous research of PprI’s crucial role in radioresistance of D. radiodurans, we have attempted to examine and compare the nucleoid morphology differences among wild-type D. ra-diodurans R1 strain, pprI function-deficient mutant (YR1), and pprI function-complementary strains (YR1001, YR1002, and YR1004) before and after exposure to ionizing irradiation. Fluorescence mi-croscopy images indicate: (1) the majority of nucleoid structures in radioresistant strain R1 cells ex-hibit the tightly packed ring-like morphology, while the pprI function-deficient mutant YR1 cells carrying predominate ring-like structure represent high sensitivity to irradiation; (2) as an extreme radioresistant strain similar to wild-type R1, pprI completely function-complementary strain YR1001 almost displays the loose and irregular nucleoid morphologies. On the other hand, another radioresistant pprI partly function-complementary strain YR1002’s nucleiods exhibit about 60% ring-like structure; (3) a PprI C-terminal deletion strain YR1004 consisting of approximately 60% of ring-like nucleoid is very sensi-tive to radiation. Therefore, our present experiments do not support the conclusion that the ring-like nucleoid of D. radiodurans does play a key role in radioresistance.
基金supported by the National Natural Science Foundation of China (Grant Nos. 30830006 and 31000045)the National High Technology Research and Development Program of China (Grant No. 2007AA021305)+3 种基金Major Scientific and Technological Project for the Creation of Significant New Drugs (Grant No. 2009ZXJ09001-034)Major Project for Genetically Modified Organism Breeding (Grant No. 2009ZX08009-075B)Application of Nuclear Techniques in Agriculture from Ministry of Agriculture of China (Grant No. 200803034) to Yue-jin Huathe 2009 Zhejiang Innovation Program for Graduates
文摘The β-subunit of RNA polymerase, which is involved in rifampin binding, is highly conserved among prokaryotes, and Rile mutants detected in many bacteria are the result of amino acid changes. Spontaneous rifampin resistance mutations resulting in amino acid replacement （L420R） and deletion （1258-66 9 bp deletion） have been previously isolated in the rpoB gene of Deinococcus radiodurans. In this study, a β-subunit mutation in D. radiodurans resulted in a unique effect on growth rate. We used DNA microarrays and biochemical assays to investigate how the Rif mutation in the β-subunit led to changes in growth rate via altered regulation of multiple genes. The expression of genes with predicted functions in metabolism, cellular processes and signaling, and information storage and processing were significantly altered in the 9 bp-deletion rpoB mutant. The consensus promoter sequence of up-regulated genes in the 9 bp-deletion rpoB mutant was identified as an AT-rich sequence. Greater levels of reactive oxygen species accumulated in the L420R and 9 bp-deletion rpoB mutants compared with wild type. These results provide insight into the molecular mechanism of how the β-subunit Rif mutation alters the regulation of multiple genes.
