{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T12:57:57Z","timestamp":1788872277549,"version":"build-2803163510"},"reference-count":47,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2015,2,24]],"date-time":"2015-02-24T00:00:00Z","timestamp":1424736000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"National Science Foundation","award":["1052188"],"award-info":[{"award-number":["1052188"]}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["HL106186"],"award-info":[{"award-number":["HL106186"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Traffic"],"published-print":{"date-parts":[[2015,4]]},"abstract":"Abstract<\/jats:title>\n \n During clathrin\u2010mediated endocytosis (\n CME<\/jats:styled-content>\n ), actin assembly provides force to drive vesicle internalization. Members of the Wiskott\u2013Aldrich syndrome protein (\n WASP<\/jats:styled-content>\n ) family play a fundamental role stimulating actin assembly.\n WASP<\/jats:styled-content>\n family proteins contain a\n WH2<\/jats:styled-content>\n motif that binds globular actin (G\u2010actin) and a central\u2010acidic motif that binds the Arp2\/3 complex, thus promoting the formation of branched actin filaments. Yeast\n WASP<\/jats:styled-content>\n (Las17) is the strongest of five factors promoting Arp2\/3\u2010dependent actin polymerization during\n CME<\/jats:styled-content>\n . It was suggested that this strong activity may be caused by a putative second G\u2010actin\u2010binding motif in Las17. Here, we describe the\n in vitro<\/jats:italic>\n and\n in vivo<\/jats:italic>\n characterization of such\n L<\/jats:italic>\n as17\n G<\/jats:italic>\n \u2010actin\u2010binding\n m<\/jats:italic>\n otif (\n LGM<\/jats:styled-content>\n ) and its dependence on a group of conserved arginine residues. Using the yeast two\u2010hybrid system,\n GST<\/jats:styled-content>\n \u2010pulldown, fluorescence polarization and pyrene\u2010actin polymerization assays, we show that\n LGM<\/jats:styled-content>\n binds G\u2010actin and is necessary for normal Arp2\/3\u2010mediated actin polymerization\n in vitro<\/jats:italic>\n . Live\u2010cell fluorescence microscopy experiments demonstrate that\n LGM<\/jats:styled-content>\n is required for normal dynamics of actin polymerization during\n CME<\/jats:styled-content>\n . Further,\n LGM<\/jats:styled-content>\n is necessary for normal dynamics of endocytic machinery components that are recruited at early, intermediate and late stages of endocytosis, as well as for optimal endocytosis of native\n CME<\/jats:styled-content>\n cargo. Both\n in vitro<\/jats:italic>\n and\n in vivo<\/jats:italic>\n experiments show that\n LGM<\/jats:styled-content>\n has relatively lower potency compared to the previously known Las17 G\u2010actin\u2010binding motif,\n WH2<\/jats:styled-content>\n . These results establish a second G\u2010actin\u2010binding motif in Las17 and advance our knowledge on the mechanism of actin assembly during\n CME<\/jats:styled-content>\n .\n <\/jats:p>\n \n \n <\/jats:p>","DOI":"10.1111\/tra.12259","type":"journal-article","created":{"date-parts":[[2015,1,23]],"date-time":"2015-01-23T07:02:55Z","timestamp":1421996575000},"page":"379-397","source":"Crossref","is-referenced-by-count":18,"title":["A Second Las17 Monomeric Actin\u2010Binding Motif Functions in Arp2\/3\u2010Dependent Actin Polymerization During Endocytosis"],"prefix":"10.1111","volume":"16","author":[{"given":"Daniel","family":"Feliciano","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Colorado State University Fort Collins CO 80523 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas O.","family":"Tolsma","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Colorado State University Fort Collins CO 80523 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristen B.","family":"Farrell","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Colorado State University Fort Collins CO 80523 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Al","family":"Aradi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Colorado State University Fort Collins CO 80523 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Santiago M.","family":"Di Pietro","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Colorado State University Fort Collins CO 80523 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,2,24]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2403"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.073395"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a016790"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2011.09.001"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.78.081307.110540"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.cellbio.17.1.517"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(03)00883-3"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2005.09.024"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2005.04.014"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2175"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.05.047"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1000604"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200708060"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.05.046"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.cellbio.19.111401.093127"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biochem-060910-094416"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2006.05.008"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0060001"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ceb.2009.01.006"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb2307"},{"key":"e_1_2_6_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrm1940"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biophys.35.040405.101936"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.77.060407.135452"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1100236108"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1100125108"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200502053"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(03)00383-X"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e11-12-1022"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.e08-09-0982"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0507021102"},{"key":"e_1_2_6_32_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.281.5373.105"},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M006407200"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2012.12.024"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2009.248"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E09-05-0429"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2009.10.062"},{"key":"e_1_2_6_38_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2010.5"},{"key":"e_1_2_6_39_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/16.16.4924"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7601646"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M402513200"},{"key":"e_1_2_6_42_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.153.1.163-168.1983"},{"key":"e_1_2_6_43_1","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/122.1.19"},{"key":"e_1_2_6_44_1","first-page":"1","article-title":"In vivo and in vitro studies of adaptor\u2010clathrin interaction","volume":"47","author":"Feliciano D","year":"2011","journal-title":"J Vis Exp"},{"key":"e_1_2_6_45_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-04-420745"},{"key":"e_1_2_6_46_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.351908"},{"key":"e_1_2_6_47_1","doi-asserted-by":"publisher","DOI":"10.1038\/35050590"},{"key":"e_1_2_6_48_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M204011200"}],"container-title":["Traffic"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ftra.12259","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ftra.12259","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/tra.12259","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,29]],"date-time":"2023-09-29T18:07:25Z","timestamp":1696010845000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/tra.12259"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,2,24]]},"references-count":47,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2015,4]]}},"alternative-id":["10.1111\/tra.12259"],"URL":"https:\/\/doi.org\/10.1111\/tra.12259","archive":["Portico"],"relation":{},"ISSN":["1398-9219","1600-0854"],"issn-type":[{"value":"1398-9219","type":"print"},{"value":"1600-0854","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,2,24]]}}}