The Essential Role of the Deinococcus radiodurans ssb Gene in Cell Survival and Radiation Tolerance
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{"title"=>"The Essential Role of the Deinococcus radiodurans ssb Gene in Cell Survival and Radiation Tolerance", "type"=>"journal", "authors"=>[{"first_name"=>"J. Scott", "last_name"=>"Lockhart", "scopus_author_id"=>"55815830500"}, {"first_name"=>"Linda C.", "last_name"=>"DeVeaux", "scopus_author_id"=>"6602231406"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84881220461", "doi"=>"10.1371/journal.pone.0071651", "sgr"=>"84881220461", "pmid"=>"23951213", "issn"=>"19326203", "pui"=>"369514814"}, "id"=>"87b992c2-7b81-3cf1-b3c1-0f1a2c06cc35", "abstract"=>"Recent evidence has implicated single-stranded DNA-binding protein (SSB) expression level as an important factor in microbial radiation resistance. The genome of the extremely radiation resistant bacterium Deinococcus radiodurans contains genes for two SSB homologs: the homodimeric, canonical Ssb, encoded by the gene ssb, and a novel pentameric protein encoded by the gene ddrB. ddrB is highly induced upon exposure to radiation, and deletions result in decreased radiation-resistance, suggesting an integral role of the protein in the extreme resistance exhibited by this organism. Although expression of ssb is also induced after irradiation, Ssb is thought to be involved primarily in replication. In this study, we demonstrate that Ssb in D. radiodurans is essential for cell survival. The lethality of an ssb deletion cannot be complemented by providing ddrB in trans. In addition, the radiation-sensitive phenotype conferred by a ddrB deletion is not alleviated by providing ssb in trans. By altering expression of the ssb gene, we also show that lower levels of transcription are required for optimal growth than are necessary for high radiation resistance. When expression is reduced to that of E. coli, ionizing radiation resistance is similarly reduced. UV resistance is also decreased under low ssb transcript levels where growth is unimpaired. These results indicate that the expression of ssb is a key component of both normal cellular metabolism as well as pathways responsible for the high radiation tolerance of D. radiodurans.", "link"=>"http://www.mendeley.com/research/essential-role-deinococcus-radiodurans-ssb-gene-cell-survival-radiation-tolerance", "reader_count"=>20, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>3, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>1, "Student > Master"=>3, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>3, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>1, "Student > Master"=>3, "Student > Bachelor"=>4, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>12, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>12}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>2}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"Japan"=>1, "Chile"=>1, "India"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1148741"], "description"=>"<p>Strains and plasmids used in this study.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology", "plasmids"], "article_id"=>768830, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.t001", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Strains_and_plasmids_used_in_this_study_/768830", "title"=>"Strains and plasmids used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148736"], "description"=>"<p>Cultures were grown in 1 mM IPTG and irradiated with 23 MeV electrons at a dose rate of 30 Gy/s. (○) GY10973/p11530 (<i>ssb<sup>+</sup></i>/empty vector); (Δ) GY10973/pSL202 (<i>ssb<sup>+</sup>/ssb</i> expression plasmid) and (◊) SL102 (Δ<i>ssb/ssb</i> expression plasmid). Appropriate dilutions were plated, and survivors counted after 3–5 days. The dashed line indicates 50% survival. Error bars indicate ± standard error.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology", "strains", "altered"], "article_id"=>768826, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.g004", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ionizing_radiation_survival_of_strains_with_altered_ssb_levels_/768826", "title"=>"Ionizing radiation survival of strains with altered <i>ssb</i> levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148730"], "description"=>"<p>Cultures initially grown in 1 mM IPTG were sub-cultured twice in either 1 mM, 0.04 mM or 0 mM IPTG. Growth in the second cycle is shown. SL102 (Δ<i>ssb/ssb</i> expression plasmid) in 1 mM (Δ), 0.04 mM (◊) and 0 mM (▾) IPTG. Since there was no difference in growth with or without IPTG of strains GY10973/p11530 (<i>ssb<sup>+</sup></i>/empty vector) (○) and GY10973/pSL202 (<i>ssb<sup>+</sup></i>/<i>ssb</i> expression plasmid) (□), only 0 mM growth is presented. Error bars indicate ± standard error.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology"], "article_id"=>768820, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.g001", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_ssb_expression_on_growth_/768820", "title"=>"Effect of <i>ssb</i> expression on growth.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148740"], "description"=>"<p>Cultures were grown in 1 mM IPTG and irradiated with 23 MeV electrons at a dose rate of 30 Gy/s. (○) SL101/p11530 (Δ<i>ddrB</i>/empty expression vector); (Δ) SL101/pSL201 (Δ<i>ddrB</i>/<i>ddrB</i> expression plasmid); and (◊) SL101/pSL202 (Δ<i>ddrB</i>/<i>ssb</i> expression plasmid). Dilutions were plated, and survivors counted after 3–5 days. The dashed line indicates 50% survival. Error bars indicate ± standard error.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology", "knockout"], "article_id"=>768829, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.g006", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ionizing_radiation_survival_of_ddrB_knockout_strains_/768829", "title"=>"Ionizing radiation survival of <i>ddrB</i> knockout strains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148739"], "description"=>"<p>GY10973/p11530 (<i>ssb<sup>+</sup></i>/empty vector), GY10973/pSL202 (<i>ssb<sup>+</sup>/ssb</i> expression plasmid) and SL102 (Δ<i>ssb/ssb</i> expression plasmid) were grown under full inducing conditions. Cultures were diluted and aliquots spotted onto plates and allowed to dry prior to exposure to UV-C. Surviving colonies were counted and used to determine surviving fractions, as described.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology", "strains", "altered"], "article_id"=>768828, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.g005", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_UV_survival_of_strains_with_altered_ssb_levels_/768828", "title"=>"UV survival of strains with altered <i>ssb</i> levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148734"], "description"=>"<p>Semi-quantitative PCR was used to compare <i>ssb</i> expression between strains. Transcript levels of <i>ssb</i> in (A) GY10973/p11530 (<i>ssb<sup>+</sup></i>/empty vector) grown in 1 mM IPTG, (B) SL102 (Δ<i>ssb/ssb</i> expression plasmid) grown in 1 mM IPTG and (C) SL102 grown in 0.04 mM IPTG are shown relative to <i>ssb</i> transcript levels in strain GY10973/pSL202 (<i>ssb</i><sup>+</sup>/<i>ss</i>b expression plasmid) grown in 1 mM IPTG. All <i>ssb</i> levels were normalized with GAPDH levels. Error bars indicate ± standard error.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology", "transcript"], "article_id"=>768824, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Relative_ssb_transcript_levels_/768824", "title"=>"Relative <i>ssb</i> transcript levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148732"], "description"=>"<p>SL102 was initially grown in 1 mM IPTG, then sub-cultured twice in either 1 mM IPTG (○) or 0 mM IPTG (Δ). Error bars indicate ± standard error.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology", "depletion"], "article_id"=>768822, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.g002", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_ssb_depletion_on_growth_/768822", "title"=>"Effect of <i>ssb</i> depletion on growth.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-09 02:08:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/1148742"], "description"=>"<p>Primers used in this study.</p>", "links"=>[], "tags"=>["astrobiology", "Extremophiles", "Biochemistry", "proteins", "DNA-binding proteins", "Protein interactions", "biophysics", "Radiation biophysics", "Radiation effects", "genetics", "gene expression", "DNA transcription", "Gene function", "microbiology", "bacteriology", "Bacterial physiology"], "article_id"=>768831, "categories"=>["Biological Sciences"], "users"=>["J. Scott Lockhart", "Linda C. DeVeaux"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0071651.t002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primers_used_in_this_study_/768831", "title"=>"Primers used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-08-09 02:08:21"}

PMC Usage Stats | Further Information

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Relative Metric

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