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ECA exists in outer membrane diacylglycerol\u2010phosphate\u2010 and lipopolysaccharide\u2010linked forms, and a cyclic periplasmic form (ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    ). Intriguingly, ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    both affects the outer membrane permeability barrier and functions in regulation of diacylglycerol\u2010phosphate\u2010linked ECA. While the length of linear ECA polymers generated by WzyE is regulated by the co\u2010polymerase WzzE, WzzE is also required for ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    biogenesis and no ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    is synthesized in its absence. To define WzzE functions necessary for ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    biosynthesis, we generated plasmid\u2010borne\n                    <jats:italic>wzzE<\/jats:italic>\n                    mutants in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                    <\/jats:styled-content>\n                    K\u201012 and quantified their effects on linear ECA regulation and ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    synthesis, determining that mutations disrupting linear ECA regulation in either transmembrane helix 2 or the periplasmic domain abolished ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    synthesis. Moreover, we identified two mutations residue F104 that differed in ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    abundance despite indistinguishable linear ECA regulation:\n                    <jats:italic>wzzE<\/jats:italic>\n                    <jats:sub>F104H<\/jats:sub>\n                    caused an approximately 2\u2010fold decrease in ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    abundance, whereas\n                    <jats:italic>wzzE<\/jats:italic>\n                    <jats:sub>F104Y<\/jats:sub>\n                    retained wild\u2010type abundance. Chromosomal\n                    <jats:italic>wzzE<\/jats:italic>\n                    mutants recapitulated this phenotype, demonstrating that these substitutions genetically uncouple levels of ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    synthesis from linear ECA regulation. Thus, although WzzE\u2010mediated chain\u2010length regulation is necessary for ECA\n                    <jats:sub>CYC<\/jats:sub>\n                    biogenesis, it is not sufficient.\n                  <\/jats:p>","DOI":"10.1111\/mmi.70088","type":"journal-article","created":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T07:04:36Z","timestamp":1781852676000},"page":"190-207","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Chain\u2010Length Regulation by\n                    <scp>WzzE<\/scp>\n                    Is Necessary for, but Genetically Separable From, Cyclic Enterobacterial Common Antigen Synthesis"],"prefix":"10.1111","volume":"126","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9228-8168","authenticated-orcid":false,"given":"Joseph F.","family":"Carr","sequence":"first","affiliation":[{"name":"Department of Biology Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yohannes H.","family":"Rezenom","sequence":"additional","affiliation":[{"name":"Department of Chemistry Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer S.","family":"Rudolf","sequence":"additional","affiliation":[{"name":"Department of Biology Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel J.","family":"Warzecha","sequence":"additional","affiliation":[{"name":"Department of Biology Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela M.","family":"Mitchell","sequence":"additional","affiliation":[{"name":"Department of Biology Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,6,19]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07487-w"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1099\/ijsem.0.001485"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00315-16"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2010.00134"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1107\/S090744499901481X"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006434"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00224-22"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/msb4100050"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)80083-7"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.181.20.6564-6568.1999"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.171.3.1326-1332.1989"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)48216-6"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1987.tb10544.x"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1128\/ecosalplus.esp-0001-2018"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2017.03.017"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002918"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.1999.01591.x"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.120163297"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-818619-0.00029-0"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.0c00983"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.185.6.1995-2004.2003"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1023\/B:JNMR.0000019252.65073.24"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.370817"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens11030356"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200351250"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.05559-11"},{"key":"e_1_2_11_28_1","first-page":"1003","article-title":"[The Structure and Significance of Enterobacterial Common Antigen (ECA)]","volume":"69","author":"Go\u017adziewicz T. 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