{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T10:59:23Z","timestamp":1758279563389},"reference-count":23,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2004,2,19]],"date-time":"2004-02-19T00:00:00Z","timestamp":1077148800000},"content-version":"vor","delay-in-days":6258,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Cellular Biochemistry"],"published-print":{"date-parts":[[1987,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The cleavage of poliovirus precursor polypeptides occurs at specific amino acid pairs that are recognized by viral proteinases. Most of the polio\u2010specific cleavages occur at glutamine\u2010glyeine (Q\u2010G) pairs that are recognized by the viral\u2010encoded proteinase 3C (formerly called P3\u20137c). In order to carry out a defined molecular genetic study of the enzymatic activity of protein 3C, we have made cDNA clones of the poliovirus genome. The cDNA region corresponding to protein 3C was inserted into an inducible bacterial expression vector. This recombinant plasmid (called pIN\u2010III\u2010C3\u20137c) utilizes the bacterial lipoprotein promoter to direct the synthesis of a precursor polypeptide that contains the amino acid sequence of protein 3C as well as the amino\u2010 and carboxy\u2010terminal Q\u2010G cleavage signals. These signals have been previously shown to allow autocatalytic production of protein 3C in bacteria transformed with plasmid pIN\u2010III\u2010C3\u20137c. We have taken advantage of the autocatalytic cleavage of 3C in a bacterial expression system to study the effects of site\u2010specific mutagenesis on its proteolytic activity. One mutation that we have introduced into the cDNA region encoding 3C is a single amino acid insertion near the carboxy\u2010terminal Q\u2010G cleavage site. The mutant recombinant plasmid (designated pIN\u2010III\u2010C3\u2010\u03bc10) directs the synthesis of a bacterial\u2010polio precursor polypeptide that is like the wild\u2010type construct (pIN\u2010III\u2010C3\u20137c). However, unlike the wild\u2010type precursor, the mutant precursor cannot undergo autocatalytic cleavage to generate the mature proteinase 3C. Rather, the precursor is able to carry out cleavage at the amino\u2010terminal Q\u2010G site but not at the carboxy\u2010terminal site. Thus, we have generated an altered poliovirus proteinase that is still able to carry out at least part of its cleavage activities but is unable to be a suitable substrate for self\u2010cleavage at its carboxy\u2010terminal Q\u2010G pair.<\/jats:p>","DOI":"10.1002\/jcb.240330105","type":"journal-article","created":{"date-parts":[[2004,12,30]],"date-time":"2004-12-30T15:18:02Z","timestamp":1104419882000},"page":"39-51","source":"Crossref","is-referenced-by-count":30,"title":["Site\u2010specific mutagenesis of cDNA clones expressing a poliovirus proteinase"],"prefix":"10.1002","volume":"33","author":[{"given":"Bert L.","family":"Semler","sequence":"first","affiliation":[]},{"given":"Victoria H.","family":"Johnson","sequence":"additional","affiliation":[]},{"given":"Patricia Gillis","family":"Dewalt","sequence":"additional","affiliation":[]},{"given":"Mary Frances","family":"Ypma\u2010Wong","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2004,2,19]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.79.13.3973"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(86)90790-7"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/291547a0"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.49.3.873-880.1984"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.78.6.3464"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(81)90242-7"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.41.1.340-344.1982"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.42.1.194-199.1982"},{"key":"e_1_2_1_10_2","unstructured":"ToyodaH NicklinMJH MurrayMG WimmerE: \u201cProtein Engineering: Application in Science Medicine and Industry.\u201d New York: Academic Press Inc. in press."},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.78.8.4887"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.78.10.5983"},{"key":"e_1_2_1_13_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/12.12.5123"},{"key":"e_1_2_1_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(84)90441-0"},{"key":"e_1_2_1_15_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/B978-0-12-372380-2.50006-8","volume-title":"Experimental Manipulation of Gene Expression","author":"Masui Y","year":"1983"},{"key":"e_1_2_1_16_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/7.6.1513"},{"key":"e_1_2_1_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0042-6822(83)80018-X"},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(73)90341-3"},{"key":"e_1_2_1_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.83.6.1777"},{"key":"e_1_2_1_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.50.3.957-959.1984"},{"key":"e_1_2_1_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.72.12.4734"},{"key":"e_1_2_1_22_2","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1128\/jvi.35.3.732-740.1980","volume":"35","author":"Van Dyke TA","year":"1980","journal-title":"J Virol"},{"key":"e_1_2_1_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.5.11.2913"},{"key":"e_1_2_1_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00665a019"}],"container-title":["Journal of Cellular Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjcb.240330105","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jcb.240330105","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,21]],"date-time":"2023-10-21T10:35:54Z","timestamp":1697884554000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jcb.240330105"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1987,1]]},"references-count":23,"journal-issue":{"issue":"1","published-print":{"date-parts":[[1987,1]]}},"alternative-id":["10.1002\/jcb.240330105"],"URL":"https:\/\/doi.org\/10.1002\/jcb.240330105","archive":["Portico"],"relation":{},"ISSN":["0730-2312","1097-4644"],"issn-type":[{"value":"0730-2312","type":"print"},{"value":"1097-4644","type":"electronic"}],"subject":[],"published":{"date-parts":[[1987,1]]}}}