{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T06:38:54Z","timestamp":1779345534924,"version":"3.51.4"},"reference-count":42,"publisher":"Proceedings of the National Academy of Sciences","issue":"27","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2008,7,8]]},"abstract":"<jats:p>\n            During translation, tRNAs must move rapidly to their adjacent sites in the ribosome while maintaining precise pairing with mRNA. This movement (translocation) occurs in a stepwise manner with hybrid-state intermediates, but it is unclear how these hybrid states relate to kinetically defined events of translocation. Here we analyze mutations at position 2394 of 23S rRNA in a pre-steady-state kinetic analysis of translocation. These mutations target the 50S E site and are predicted to inhibit P\/E state formation. Each mutation decreases growth rate, the maximal rate of translocation (\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>trans<\/jats:sub>\n            ), and the apparent affinity of EF-G for the pretranslocation complex (i.e., increases\n            <jats:italic>K<\/jats:italic>\n            <jats:sub>1\/2<\/jats:sub>\n            ). The magnitude of these defects follows the trend A &gt; G &gt; U. Because the C2394A mutation did not decrease the rate of single-turnover GTP hydrolysis, the &gt;20-fold increase in\n            <jats:italic>K<\/jats:italic>\n            <jats:sub>1\/2<\/jats:sub>\n            conferred by C2394A can be attributed to neither the initial binding of EF-G nor the subsequent GTP hydrolysis step. We propose that C2394A inhibits a later step, P\/E state formation, to confer its effects on translocation. Replacement of the peptidyl group with an aminoacyl group, which is predicted to inhibit A\/P state formation, decreases\n            <jats:italic>k<\/jats:italic>\n            <jats:sub>trans<\/jats:sub>\n            without increasing\n            <jats:italic>K<\/jats:italic>\n            <jats:sub>1\/2<\/jats:sub>\n            . These data suggest that movements of tRNA into the P\/E and A\/P sites are separable events. This mutational study allows tRNA movements with respect to both subunits to be integrated into a kinetic model for translocation.\n          <\/jats:p>","DOI":"10.1073\/pnas.0710146105","type":"journal-article","created":{"date-parts":[[2008,7,1]],"date-time":"2008-07-01T01:10:47Z","timestamp":1214874647000},"page":"9192-9197","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":71,"title":["Role of hybrid tRNA-binding states in ribosomal translocation"],"prefix":"10.1073","volume":"105","author":[{"given":"Sarah E.","family":"Walker","sequence":"first","affiliation":[{"name":"*Department of Microbiology and"}],"role":[{"role":"author","vocab":"crossref"}]},{"given":"Shinichiro","family":"Shoji","sequence":"additional","affiliation":[{"name":"*Department of Microbiology and"}],"role":[{"role":"author","vocab":"crossref"}]},{"given":"Dongli","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104"}],"role":[{"role":"author","vocab":"crossref"}]},{"given":"Barry S.","family":"Cooperman","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104"}],"role":[{"role":"author","vocab":"crossref"}]},{"given":"Kurt","family":"Fredrick","sequence":"additional","affiliation":[{"name":"*Department of Microbiology and"},{"name":"Ohio State Biochemistry Program, Ohio State University, Columbus, OH 43210; and"}],"role":[{"role":"author","vocab":"crossref"}]}],"member":"341","published-online":{"date-parts":[[2008,7,8]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1038\/342142a0"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00500a004"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0403884101"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.01.022"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2005.10.007"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(03)00476-8"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0401904101"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7601677"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0079-6603(08)60346-3"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/385037a0"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.01.014"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(03)00230-2"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2004.08.097"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7600884"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1989.tb08574.x"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.211184098"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb1060"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1017\/S1355838201001650"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1261\/rna.5120503"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.08.032"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1131127"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gki910"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(03)00146-3"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/80938"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(02)00523-3"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb1074"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1261\/rna.601507"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/10695"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/35018597"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2004.01.013"},{"key":"e_1_3_3_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2007.04.042"},{"key":"e_1_3_3_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(03)00478-1"},{"key":"e_1_3_3_33_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0706784104"},{"key":"e_1_3_3_34_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2005.09.008"},{"key":"e_1_3_3_35_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00671a020"},{"key":"e_1_3_3_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)40211-1"},{"key":"e_1_3_3_37_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.131549798"},{"key":"e_1_3_3_38_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi027316z"},{"key":"e_1_3_3_39_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M405603200"},{"key":"e_1_3_3_40_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi0516677"},{"key":"e_1_3_3_41_2","doi-asserted-by":"publisher","DOI":"10.1261\/rna.715307"},{"key":"e_1_3_3_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2006.05.006"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.0710146105","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T20:31:52Z","timestamp":1649795512000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.0710146105"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,7,8]]},"references-count":42,"journal-issue":{"issue":"27","published-print":{"date-parts":[[2008,7,8]]}},"alternative-id":["10.1073\/pnas.0710146105"],"URL":"https:\/\/doi.org\/10.1073\/pnas.0710146105","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,7,8]]},"assertion":[{"value":"2007-10-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-07-08","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}