{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T05:05:03Z","timestamp":1774069503273,"version":"3.50.1"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2014,3,1]],"date-time":"2014-03-01T00:00:00Z","timestamp":1393632000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2014,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Mycobacteria contain genes for several DNA-dependent RNA primases, including\n            <jats:italic>dnaG<\/jats:italic>\n            , which encodes an essential replication enzyme that has been proposed as a target for antituberculosis compounds. An\n            <jats:italic>in silico<\/jats:italic>\n            analysis revealed that mycobacteria also possess archaeo-eukaryotic superfamily primases (AEPs) of unknown function. Using a homologous recombination system, we obtained direct evidence that wild-type\n            <jats:italic>dnaG<\/jats:italic>\n            cannot be deleted from the chromosome of\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Mycobacterium smegmatis<\/jats:named-content>\n            without disrupting viability, even in backgrounds in which mycobacterial AEPs are overexpressed. In contrast, single-deletion AEP mutants or mutants defective for all four identified\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">M. smegmatis<\/jats:named-content>\n            AEP genes did not exhibit growth defects under standard laboratory conditions. Deletion of native\n            <jats:italic>dnaG<\/jats:italic>\n            in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">M. smegmatis<\/jats:named-content>\n            was tolerated only after the integration of an extra intact copy of the\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">M. smegmatis<\/jats:named-content>\n            or\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Mycobacterium tuberculosis<\/jats:named-content>\n            <jats:italic>dnaG<\/jats:italic>\n            gene, under the control of chemically inducible promoters, into the\n            <jats:italic>attB<\/jats:italic>\n            site of the chromosome.\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">M. tuberculosis<\/jats:named-content>\n            and\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">M. smegmatis<\/jats:named-content>\n            DnaG proteins were overproduced and purified, and their primase activities were confirmed using radioactive RNA synthesis assays. The enzymes appeared to be sensitive to known inhibitors (suramin and doxorubicin) of DnaG. Notably,\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">M. smegmatis<\/jats:named-content>\n            bacilli appeared to be sensitive to doxorubicin and resistant to suramin. The growth and survival of conditional mutant mycobacterial strains in which DnaG was significantly depleted were only slightly affected under standard laboratory conditions. Thus, although DnaG is essential for mycobacterial viability, only low levels of protein are required for growth. This suggests that very efficient inhibition of enzyme activity would be required for mycobacterial DnaG to be useful as an antibiotic target.\n          <\/jats:p>","DOI":"10.1128\/aac.01721-13","type":"journal-article","created":{"date-parts":[[2013,12,31]],"date-time":"2013-12-31T02:28:14Z","timestamp":1388456894000},"page":"1699-1706","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":20,"title":["Evaluation of DNA Primase DnaG as a Potential Target for Antibiotics"],"prefix":"10.1128","volume":"58","author":[{"given":"Aneta","family":"Kuron","sequence":"first","affiliation":[{"name":"Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Malgorzata","family":"Korycka-Machala","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Brzostek","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marcin","family":"Nowosielski","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aidan","family":"Doherty","sequence":"additional","affiliation":[{"name":"Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bozena","family":"Dziadek","sequence":"additional","affiliation":[{"name":"Department of Immunoparasitology, University of Lodz, Lodz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jaroslaw","family":"Dziadek","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Polish Academy of Sciences, Lodz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1185449"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6976.2012.00331.x"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1378\/chest.08-2427"},{"key":"e_1_3_3_5_2","first-page":"301","article-title":"Emergence of Mycobacterium tuberculosis with extensive resistance to second-line drugs worldwide, 2000-2004. 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