A DNA Damage-Induced, SOS-Independent Checkpoint Regulates Cell Division in Caulobacter crescentus
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{"title"=>"A DNA Damage-Induced, SOS-Independent Checkpoint Regulates Cell Division in Caulobacter crescentus", "type"=>"journal", "authors"=>[{"first_name"=>"Joshua W.", "last_name"=>"Modell", "scopus_author_id"=>"57197429592"}, {"first_name"=>"Tracy K.", "last_name"=>"Kambara", "scopus_author_id"=>"56468922200"}, {"first_name"=>"Barrett S.", "last_name"=>"Perchuk", "scopus_author_id"=>"8889357900"}, {"first_name"=>"Michael T.", "last_name"=>"Laub", "scopus_author_id"=>"7006013323"}], "year"=>2014, "source"=>"PLoS Biology", "identifiers"=>{"issn"=>"15457885", "scopus"=>"2-s2.0-84920468768", "pui"=>"601112631", "doi"=>"10.1371/journal.pbio.1001977", "isbn"=>"1545-7885", "sgr"=>"84920468768", "pmid"=>"25350732"}, "id"=>"69e81258-b1b0-3d03-a066-e60c6719a68b", "abstract"=>"Cells must coordinate DNA replication with cell division, especially during episodes of DNA damage. The paradigm for cell division control following DNA damage in bacteria involves the SOS response where cleavage of the transcriptional repressor LexA induces a division inhibitor. However, in Caulobacter crescentus, cells lacking the primary SOS-regulated inhibitor, sidA, can often still delay division post-damage. Here we identify didA, a second cell division inhibitor that is induced by DNA damage, but in an SOS-independent manner. Together, DidA and SidA inhibit division, such that cells lacking both inhibitors divide prematurely following DNA damage, with lethal consequences. We show that DidA does not disrupt assembly of the division machinery and instead binds the essential division protein FtsN to block cytokinesis. Intriguingly, mutations in FtsW and FtsI, which drive the synthesis of septal cell wall material, can suppress the activity of both SidA and DidA, likely by causing the FtsW/I/N complex to hyperactively initiate cell division. Finally, we identify a transcription factor, DriD, that drives the SOS-independent transcription of didA following DNA damage.", "link"=>"http://www.mendeley.com/research/dna-damageinduced-sosindependent-checkpoint-regulates-cell-division-caulobacter-crescentus", "reader_count"=>48, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>8, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>1, "Student > Master"=>4, "Student > Bachelor"=>10, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Librarian"=>1, "Researcher"=>8, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>1, "Student > Master"=>4, "Student > Bachelor"=>10, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>11, "Agricultural and Biological Sciences"=>30, "Medicine and Dentistry"=>2, "Physics and Astronomy"=>1, "Chemistry"=>1, "Social Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Chemistry"=>{"Chemistry"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>30}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>11}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Belgium"=>1, "United States"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1767296"], "description"=>"<p>Growth curves (A) and micrographs (B) of strains overexpressing <i>didA</i>. Cells harboring a low- or medium-copy plasmid that expresses <i>didA</i> from the vanillate-inducible promoter P<i><sub>van</sub></i> were grown in rich medium with or without vanillate for the times indicated. Bar, 2 µm.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220542, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g002", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DidA_is_sufficient_to_inhibit_cell_division_/1220542", "title"=>"DidA is sufficient to inhibit cell division.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767497", "https://ndownloader.figshare.com/files/1767498", "https://ndownloader.figshare.com/files/1767499", "https://ndownloader.figshare.com/files/1767503", "https://ndownloader.figshare.com/files/1767504", "https://ndownloader.figshare.com/files/1767506", "https://ndownloader.figshare.com/files/1767507", "https://ndownloader.figshare.com/files/1767508", "https://ndownloader.figshare.com/files/1767509", "https://ndownloader.figshare.com/files/1767510", "https://ndownloader.figshare.com/files/1767511", "https://ndownloader.figshare.com/files/1767512", "https://ndownloader.figshare.com/files/1767514", "https://ndownloader.figshare.com/files/1767515", "https://ndownloader.figshare.com/files/1767516"], "description"=>"<div><p>Cells must coordinate DNA replication with cell division, especially during episodes of DNA damage. The paradigm for cell division control following DNA damage in bacteria involves the SOS response where cleavage of the transcriptional repressor LexA induces a division inhibitor. However, in <i>Caulobacter crescentus</i>, cells lacking the primary SOS-regulated inhibitor, <i>sidA</i>, can often still delay division post-damage. Here we identify <i>didA</i>, a second cell division inhibitor that is induced by DNA damage, but in an SOS-independent manner. Together, DidA and SidA inhibit division, such that cells lacking both inhibitors divide prematurely following DNA damage, with lethal consequences. We show that DidA does not disrupt assembly of the division machinery and instead binds the essential division protein FtsN to block cytokinesis. Intriguingly, mutations in FtsW and FtsI, which drive the synthesis of septal cell wall material, can suppress the activity of both SidA and DidA, likely by causing the FtsW/I/N complex to hyperactively initiate cell division. Finally, we identify a transcription factor, DriD, that drives the SOS-independent transcription of <i>didA</i> following DNA damage.</p></div>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220617, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>["https://dx.doi.org/10.1371/journal.pbio.1001977.s001", "https://dx.doi.org/10.1371/journal.pbio.1001977.s002", "https://dx.doi.org/10.1371/journal.pbio.1001977.s003", "https://dx.doi.org/10.1371/journal.pbio.1001977.s004", "https://dx.doi.org/10.1371/journal.pbio.1001977.s005", "https://dx.doi.org/10.1371/journal.pbio.1001977.s006", "https://dx.doi.org/10.1371/journal.pbio.1001977.s007", "https://dx.doi.org/10.1371/journal.pbio.1001977.s008", "https://dx.doi.org/10.1371/journal.pbio.1001977.s009", "https://dx.doi.org/10.1371/journal.pbio.1001977.s010", "https://dx.doi.org/10.1371/journal.pbio.1001977.s011", "https://dx.doi.org/10.1371/journal.pbio.1001977.s012", "https://dx.doi.org/10.1371/journal.pbio.1001977.s013", "https://dx.doi.org/10.1371/journal.pbio.1001977.s014", "https://dx.doi.org/10.1371/journal.pbio.1001977.s015"], "stats"=>{"downloads"=>19, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_DNA_Damage_Induced_SOS_Independent_Checkpoint_Regulates_Cell_Division_in_Caulobacter_crescentus_/1220617", "title"=>"A DNA Damage-Induced, SOS-Independent Checkpoint Regulates Cell Division in <i>Caulobacter crescentus</i>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767359"], "description"=>"<p>(A) Schematic showing the membrane topology of FtsW, FtsI, FtsN, SidA, and DidA. Missense mutations and the GFP-FtsN fusion that suppress the activities of SidA or DidA, or both, are listed in red. (B) Strains harboring the mutations indicated were transformed with a medium-copy plasmid expressing <i>M2-sidA</i> from the P<i><sub>xyl</sub></i> promoter or a low-copy plasmid expressing <i>3×M2-didA</i> from the P<i><sub>lac</sub></i> promoter. To induce <i>M2-sidA</i>, strains were grown in media supplemented with glucose and then plated on media supplemented with xylose. To induce <i>3×M2-didA</i>, strains were grown in media without inducer and then plated with IPTG. Each strain was plated in 10-fold dilutions. (C) Bacterial two-hybrid analysis of interactions between T25-DidA or T25-FtsW and T18 fusions to FtsN and the mutants indicated. Below is a graphical representation of each T18 construct. (D) Strains harboring the mutations indicated were transformed with plasmids for inducing M2-SidA or DidA and plated on inducing media.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220564, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g005", "stats"=>{"downloads"=>5, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutations_in_the_FtsW_FtsI_FtsN_complex_suppress_SidA_and_DidA_overproduction_phenotypes_/1220564", "title"=>"Mutations in the FtsW/FtsI/FtsN complex suppress SidA and DidA overproduction phenotypes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767382"], "description"=>"<p>(A–B) Two cell division inhibitors are induced following DNA damage in <i>Caulobacter</i>. <i>sidA</i> is induced by cleavage of the SOS repressor LexA while <i>didA</i> is induced by DriD. SidA and DidA are small transmembrane proteins that can block cell division by preventing the divisome subcomplex FtsW/I/N from assuming an active state, designated FtsW/I/N*. FtsW/I/N* could promote division by enhancing peptidoglycan synthesis and remodeling, by triggering FtsZ constriction, or by coordinating these activities.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220574, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g008", "stats"=>{"downloads"=>3, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Two_independent_pathways_regulate_cell_division_in_Caulobacter_following_DNA_damage_/1220574", "title"=>"Two independent pathways regulate cell division in <i>Caulobacter</i> following DNA damage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767345"], "description"=>"<p>(A) The subcellular localization of DidA was examined in a strain expressing <i>M2-yfp-didA</i> from the xylose-inducible promoter P<i><sub>xyl</sub></i> on a low-copy plasmid. Cells were grown to mid-exponential phase in rich media with glucose and then shifted to xylose. At the times indicated, cells were imaged by phase and epifluorescent microscopy. In the fluorescent micrographs, cell boundaries were added after imaging. (B) Subcellular fractionation of cells overexpressing <i>3×M2-didA</i> from the P<i><sub>van</sub></i> promoter on a medium-copy plasmid for 1.5 hours and expressing the transmembrane protein <i>cckA-gfp</i> from P<i><sub>cckA</sub></i> on the chromosome. Samples were fractionated into soluble (S) and membrane (M) fractions and analyzed by Western blot. The membrane was cut into three pieces, indicated by dashed lines, and probed with antibodies specific for the GFP, CtrA, or M2 epitope. (C) Bacterial two-hybrid analysis of interactions between T25-DidA and cell division proteins fused to T18, as indicated. The FtsIΔC construct lacking the C-terminal catalytic domain previously showed interactions with FtsW and FtsN as expected, unlike the full-length version of FtsI <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977-Modell1\" target=\"_blank\">[13]</a>. The interacting pair T18-M2-SidA and T25-FtsN was included for comparison. <i>E. coli</i> strains harboring each pair of fusions were plated on LB, and colonies were restruck on MacConkey plates containing maltose. Red streaks indicate positive interactions. −/− indicates empty vectors negative control, +/+ indicates the zip/zip fusions used as a positive control. (D) Subcellular localization of FtsZ, FtsW, FtsI, and FtsN were examined in strains expressing <i>ftsZ-yfp</i> from the chromosomal P<i><sub>van</sub></i> promoter, or <i>venus-ftsW</i>, <i>gfp-ftsI</i> or <i>gfp-ftsN</i> from its native chromosomal locus. Each strain was transformed with a medium-copy plasmid expressing <i>didA</i> from the P<i><sub>van</sub></i> promoter. Strains were grown to mid-exponential phase and samples imaged by phase and epifluorescent microscopy after addition of vanillate for 4.5 hours. In the fluorescent images, cell outlines were drawn based on the phase micrographs. Bar, 2 µm. (E) Strains from (D) were grown to mid-exponential phase and 10-fold serial dilutions were plated on rich media supplemented with vanillate to induce <i>didA</i> expression.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220558, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g004", "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DidA_is_a_small_inner_membrane_protein_that_interacts_with_FtsN_/1220558", "title"=>"DidA is a small, inner membrane protein that interacts with FtsN.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767372"], "description"=>"<p>(A) Wild-type cells harboring low-copy plasmids expressing <i>egfp</i> from the <i>sidA</i> or <i>didA</i> promoters were treated with MMC (0.5 and 3 µg/ml), hydroxyurea (HU; 0.5 and 3 mg/ml) or zeocin (2.5 and 15 µg/ml) and then analyzed by Western blot using an α-GFP antibody. (B) Diagram of <i>driD</i> indicating the predicted helix-turn-helix (HTH) and WYL domains. Arrows indicate transposon insertion sites in the genetic screen that identified <i>driD</i>. (C) Wild-type, <i>ΔdriD</i>, and <i>ΔrecA</i> cells were transformed with the P<i><sub>sidA</sub></i> and P<i><sub>didA</sub></i> reporter plasmids from (A) and treated with 3 µg/ml MMC or 15 µg/ml zeocin for 1 hour. Samples were analyzed by Western blot using an α-GFP antibody. (D) 10-fold serial dilutions of the strains indicated were grown on plates containing 0.35 µg/ml MMC. (E) <i>ΔdriD</i> cells carrying a low-copy plasmid producing a control construct (P<i><sub>xyl</sub>-ftsW-egfp</i>), untagged DidA, or DidA fused at either its N- or C-terminal end to a 3×M2 tag and expressed from the <i>didA</i> promoter were treated with 15 µg/ml zeocin for 45 minutes. Samples were analyzed by Western blot using an α-FLAG/M2 antibody. (F) <i>ΔdriD</i> cells carrying a low-copy plasmid expressing either <i>driD</i> or <i>driD-3×M2</i> from the <i>driD</i> promoter were treated with 15 µg/ml zeocin for 45 minutes. DriD was immunoprecipitated with an α-FLAG/M2 antibody and promoter occupancy was analyzed by quantitative PCR using primers specific for P<i><sub>didA</sub></i>. Fold-enrichment values were normalized relative to the enrichment of a region within the coding sequence of <i>ruvA</i>. For raw data, see <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977.s014\" target=\"_blank\">Data S3</a>.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220569, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g007", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DriD_directly_activates_didA_/1220569", "title"=>"DriD directly activates <i>didA</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767289"], "description"=>"<p>(A) Wild-type and Δ<i>recA</i> cells were grown in rich medium to mid-exponential phase and treated with 1 µg/ml MMC for 30 minutes. Expression values, the average of two biological replicates, are shown for the 50 most upregulated genes in wild-type cells with fold-change ratios calculated in comparison to mock treated cells. The dashed line corresponds to fold-change values that are identical in wild-type and <i>ΔrecA</i> cells. For complete data, see <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977.s002\" target=\"_blank\">Figure S2</a> and <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977.s012\" target=\"_blank\">Data S1A</a>. (B) CC3114 and CCNA03212 (<i>didA</i>) are shown schematically in their genomic context. Nucleotide positions relative to the annotated CC3114 start site are shown below. The gray shaded region represents a predicted transmembrane domain. (C) Western blot of cells producing DidA fused to a C-terminal 3×M2 epitope from the chromosomal <i>didA</i> locus. Cells were grown to mid-exponential phase and treated with 1 µg/ml MMC for the times indicated. (D) Western blot of wild-type, Δ<i>recA</i> and <i>lexA(K203A)</i> cells expressing <i>didA-3×M2</i> from its native locus treated with 1 µg/ml MMC for 1 hour. Membranes (C–D) were blotted with the α-FLAG/M2 antibody.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220535, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g001", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_didA_is_induced_by_DNA_damage_and_is_not_SOS_regulated_/1220535", "title"=>"<i>didA</i> is induced by DNA damage and is not SOS regulated.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767369"], "description"=>"<p>(A) The strains indicated were grown to mid-exponential phase in rich media and imaged by phase microscopy. Bar, 2 µm. (B) Each strain indicated was grown to mid-exponential phase and average cell length, relative to wild-type, was calculated (all <i>n</i>>440). Error bars represent standard error of the mean (SEM), and asterisks indicate <i>p</i><0.01 (*) or <i>p</i><0.0001 (**). The strain denoted <i>ftsW**I*</i> combines the mutations <i>ftsW(F145L, A246T)</i> and <i>ftsI(I45V)</i>. Separate graphs are shown for cell length measurements made on different days. For raw data, see <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977.s014\" target=\"_blank\">Data S3</a>. (C) Wild-type, <i>ΔsidAΔdidA</i>, <i>ftsW(A246T)</i>, and <i>ftsW**I*</i> cells were grown to mid-exponential phase and plated in 10-fold dilutions on rich media containing no additives, 0.35 µg/ml MMC or 6 µg/ml cephalexin. (D) The strains from (C) were grown to mid-exponential phase in rich media and treated with MMC or cephalexin at the concentrations in (C) for 6 hours. PI at 5 µM was added 1.5 hours before imaging. Cells were imaged by phase and fluorescence microscopy; cell lengths and percentage of PI+ cells are shown by bar graphs. For raw data, see <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977.s014\" target=\"_blank\">Data S3</a>.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220568, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g006", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutations_that_suppress_sidA_and_didA_overexpression_likely_hyperactivate_cell_division_/1220568", "title"=>"Mutations that suppress <i>sidA</i> and <i>didA</i> overexpression likely hyperactivate cell division.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/1767308"], "description"=>"<p>(A) Wild-type, <i>ΔsidA</i>, <i>ΔdidA</i>, and <i>ΔsidAΔdidA</i> cells were grown to mid-exponential phase and plated in 10-fold dilutions on rich media with or without 0.35 µg/ml MMC. (B) Wild-type and <i>ΔsidAΔdidA</i> cells carrying an empty plasmid, and <i>ΔsidAΔdidA</i> cells carrying a plasmid with either <i>sidA</i> or <i>didA</i> driven by its native promoter were plated as in (A). (C–E) Synchronous populations of swarmer cells from the strains in (A) were placed on agarose pads containing rich media and MMC and imaged for 8 hours by time-lapse microscopy. (C) The time to first mid-cell division and (D) the percentage of cells that stopped growing following division relative to the wild type are shown (for criteria on calling divisions and growth cessation, see <a href=\"http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001977#pbio.1001977.s015\" target=\"_blank\">Text S1</a>). The data in (C) are representative of biological duplicates. The data in (D) are averaged from biological duplicates. Asterisks represent a statistically significant (<i>p</i><0.01) difference relative to the wild type. Error bars represent standard error of the mean (SEM). (E) Representative fields of wild-type and <i>ΔsidAΔdidA</i> swarmer cells grown on pads containing MMC at the time points indicated in hours. Black arrows indicate cells that divided. Gray arrows indicate cells arrested for growth following division. Bar, 2 µm.</p>", "links"=>[], "tags"=>["transcriptional repressor LexA", "cell division control", "cell division", "division protein FtsN", "sos", "Caulobacter crescentus Cells", "DNA damage", "cell division inhibitor", "septal cell wall material"], "article_id"=>1220548, "categories"=>["Uncategorised"], "users"=>["Joshua W. Modell", "Tracy K. Kambara", "Barrett S. Perchuk", "Michael T. Laub"], "doi"=>"https://dx.doi.org/10.1371/journal.pbio.1001977.g003", "stats"=>{"downloads"=>4, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cells_lacking_sidA_and_didA_cannot_properly_regulate_cell_division_following_DNA_damage_/1220548", "title"=>"Cells lacking <i>sidA</i> and <i>didA</i> cannot properly regulate cell division following DNA damage.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-10-28 18:53:36"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Physical sciences/Physics", "average_usage"=>[266]}]}
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