{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T12:09:14Z","timestamp":1789733354247,"version":"4.0.1"},"reference-count":47,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2002,10,24]],"date-time":"2002-10-24T00:00:00Z","timestamp":1035417600000},"content-version":"vor","delay-in-days":23,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2002,10]]},"abstract":"<jats:title>Summary<\/jats:title>\n                  <jats:p>\n                    A putative PEST sequence was recently identified close to the N\u2010terminus of listeriolysin O (LLO), a major virulence factor secreted by the pathogenic\n                    <jats:italic>Listeria monocytogenes<\/jats:italic>\n                    . The deletion of this motif did not affect the secretion and haemolytic activity of LLO, but abolished bacterial virulence. Here, we first tested whether the replacement of the PEST motif of LLO by two different sequences, with either a very high or no PEST score, would affect phagosomal escape, protein stability and, ultimately, the virulence of\n                    <jats:italic>L. monocytogenes<\/jats:italic>\n                    . Then, we constructed LLO mutants with an intact PEST sequence but carrying mutations     on     either     side,     or     on     both     sides,     of the PEST motif. The properties of these mutants prompted us to construct three LLO mutants carrying single amino acid substitutions in the distal portion of the PEST region (P49A, K50A and P52A; preprotein numbering). Our data demonstrate that the susceptibility  of  LLO  to  intracellular  proteolytic  degradation is not related to the presence of a high PEST score sequence and that the insertion of two residues immediately downstream of the intact PEST sequence is sufficient to impair phagosomal escape and abolish bacterial virulence\n                    <jats:italic>.<\/jats:italic>\n                    Furthermore, we show that single amino acid substitutions in the distal portion of the PEST motif are sufficient to attenuate bacterial \u00advirulence   significantly,   unravelling   the   critical   role of this region of LLO in the pathogenesis of\n                    <jats:italic>L. \u00admonocytogenes<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1046\/j.1365-2958.2002.03176.x","type":"journal-article","created":{"date-parts":[[2003,3,12]],"date-time":"2003-03-12T06:46:03Z","timestamp":1047451563000},"page":"367-379","source":"Crossref","is-referenced-by-count":45,"title":["Critical role of the N\u2010terminal residues of listeriolysin O in phagosomal escape and virulence of\n                    <i>Listeria monocytogenes<\/i>"],"prefix":"10.1111","volume":"46","author":[{"given":"Marie\u2010Annick","family":"Lety","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claude","family":"Frehel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Berche","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Charbit","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2002,10,24]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1438-4221(00)80039-9"},{"key":"e_1_2_6_3_1","volume-title":"Current Protocols in Molecular Biology","author":"Ausubel 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