{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T11:25:06Z","timestamp":1772796306509,"version":"3.50.1"},"reference-count":42,"publisher":"Proceedings of the National Academy of Sciences","issue":"26","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1997,12,23]]},"abstract":"<jats:p>\n            Initiation and control of replication of the broad-host-range plasmid RK2 requires two plasmid-encoded elements, the replication origin (\n            <jats:italic>oriV<\/jats:italic>\n            ) and the initiation protein TrfA. Purified TrfA is largely in the form of a dimer; however, only the monomeric form of the protein can bind specifically to the direct repeats (iterons) at the RK2 origin. The largely dimeric form of wild-type TrfA is inactive in the initiation of replication of RK2 in an\n            <jats:italic>in vitro<\/jats:italic>\n            replication system reconstituted from purified components. However, preincubation of the TrfA protein with the ClpX molecular chaperone isolated from\n            <jats:italic>Escherichia coli<\/jats:italic>\n            activates the initiator protein for replication in the purified system. We further observed that ClpX, in an ATP-dependent reaction, greatly increases the proportion of TrfA monomers and, therefore, the ability of this protein to bind to iterons localized within RK2 origin. Finally, a copy-up mutant of the TrfA protein which is largely in the monomer form is active in the reconstituted\n            <jats:italic>in vitro<\/jats:italic>\n            replication system, and its activity is not affected by ClpX.\n          <\/jats:p>","DOI":"10.1073\/pnas.94.26.14378","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T14:42:40Z","timestamp":1027694560000},"page":"14378-14382","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":52,"title":["The replication initiation protein of the broad-host-range plasmid RK2 is activated by the ClpX\u2009chaperone"],"prefix":"10.1073","volume":"94","author":[{"given":"Igor","family":"Konieczny","sequence":"first","affiliation":[{"name":"Department of Biology, Center for Molecular Genetics, University of California, San Diego, La Jolla, CA 92093-0634; and Department of Molecular and Cellular Biology, University of Gdansk, 24 Kladki, 80-822 Gdansk, Poland"}]},{"given":"Donald R.","family":"Helinski","sequence":"additional","affiliation":[{"name":"Department of Biology, Center for Molecular Genetics, University of California, San Diego, La Jolla, CA 92093-0634; and Department of Molecular and Cellular Biology, University of Gdansk, 24 Kladki, 80-822 Gdansk, Poland"}]}],"member":"341","published-online":{"date-parts":[[1997,12,23]]},"reference":[{"key":"e_1_3_2_1_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.18.6632"},{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.21.10345"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.175.11.3546-3555.1993"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00260494"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.25.12218"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)41572-4"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1995.tb07179.x"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.9.19.2399"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1996.tb00428.x"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1996.421404.x"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.8.3274"},{"key":"e_1_3_2_12_2","first-page":"81","volume":"17","author":"Chattoraj D K","year":"1995","unstructured":"D K Chattoraj Genet Eng NY 17, 81\u201398 (1995).","journal-title":"Genet Eng NY"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.19.8923"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.9.3839"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.18.7903"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.12.7072"},{"key":"e_1_3_2_17_2","first-page":"1","volume-title":"Promiscuous Plasmids of Gram-Negative Bacteria","author":"Thomas C M","year":"1989","unstructured":"C M Thomas, D R Helinski Promiscuous Plasmids of Gram-Negative Bacteria, ed C M Thomas (Academic Press, San Diego, CA), pp. 1\u201325 (1989)."},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.76.4.1648"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.141.1.213-222.1980"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.164.1.446-455.1985"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.169.10.4703-4709.1987"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-2836(88)90118-0"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.4.1389"},{"key":"e_1_3_2_24_2","unstructured":"Konieczny I. & Helinski D. 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