{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T20:29:03Z","timestamp":1772137743530,"version":"3.50.1"},"reference-count":39,"publisher":"Rockefeller University Press","issue":"4","license":[{"start":{"date-parts":[[2020,3,25]],"date-time":"2020-03-25T00:00:00Z","timestamp":1585094400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000781","name":"European Research Council","doi-asserted-by":"publisher","award":["281971"],"award-info":[{"award-number":["281971"]}],"id":[{"id":"10.13039\/501100000781","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["WT095829AIA"],"award-info":[{"award-number":["WT095829AIA"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","award":["N000846\/1"],"award-info":[{"award-number":["N000846\/1"]}],"id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["108467\/Z\/15\/Z"],"award-info":[{"award-number":["108467\/Z\/15\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["092096"],"award-info":[{"award-number":["092096"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000289","name":"Cancer Research UK","doi-asserted-by":"publisher","award":["C6946\/A14492"],"award-info":[{"award-number":["C6946\/A14492"]}],"id":[{"id":"10.13039\/501100000289","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["105602\/Z\/14\/Z"],"award-info":[{"award-number":["105602\/Z\/14\/Z"]}],"id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Herchel Smith"},{"name":"Funai Foundation"}],"content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2020,4,6]]},"abstract":"<jats:p>Filopodia are finger-like actin-rich protrusions that extend from the cell surface and are important for cell\u2013cell communication and pathogen internalization. The small size and transient nature of filopodia combined with shared usage of actin regulators within cells confounds attempts to identify filopodial proteins. Here, we used phage display phenotypic screening to isolate antibodies that alter the actin morphology of filopodia-like structures (FLS) in vitro. We found that all of the antibodies that cause shorter FLS interact with SNX9, an actin regulator that binds phosphoinositides during endocytosis and at invadopodia. In cells, we discover SNX9 at specialized filopodia in Xenopus development and that SNX9 is an endogenous component of filopodia that are hijacked by Chlamydia entry. We show the use of antibody technology to identify proteins used in filopodia-like structures, and a role for SNX9 in filopodia.<\/jats:p>","DOI":"10.1083\/jcb.201909178","type":"journal-article","created":{"date-parts":[[2020,2,1]],"date-time":"2020-02-01T02:21:45Z","timestamp":1580523705000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":14,"title":["A direct role for SNX9 in the biogenesis of filopodia"],"prefix":"10.1083","volume":"219","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3218-1742","authenticated-orcid":true,"given":"Iris K.","family":"Jarsch","sequence":"first","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7986-4917","authenticated-orcid":true,"given":"Jonathan R.","family":"Gadsby","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annalisa","family":"Nuccitelli","sequence":"additional","affiliation":[{"name":"Antibody Discovery and Protein Engineering, AstraZeneca, Granta Park, Cambridge, UK 2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia","family":"Mason","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanae","family":"Shimo","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ludovic","family":"Pilloux","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Cambridge, Cambridge, UK 3"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bishara","family":"Marzook","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2989-2052","authenticated-orcid":true,"given":"Claire M.","family":"Mulvey","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9363-654X","authenticated-orcid":true,"given":"Ulrich","family":"Dobramysl","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3528-458X","authenticated-orcid":true,"given":"Charles R.","family":"Bradshaw","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kathryn S.","family":"Lilley","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Cambridge, Cambridge, UK 4"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard D.","family":"Hayward","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Cambridge, Cambridge, UK 3"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3652-8232","authenticated-orcid":true,"given":"Tristan J.","family":"Vaughan","sequence":"additional","affiliation":[{"name":"Antibody Discovery and Protein Engineering, AstraZeneca, Granta Park, Cambridge, UK 2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claire L.","family":"Dobson","sequence":"additional","affiliation":[{"name":"Antibody Discovery and Protein Engineering, AstraZeneca, Granta Park, Cambridge, UK 2"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9978-1382","authenticated-orcid":true,"given":"Jennifer L.","family":"Gallop","sequence":"additional","affiliation":[{"name":"Gurdon Institute and Department of Biochemistry, University of Cambridge, Cambridge, UK 1"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[2020,3,25]]},"reference":[{"key":"2023072811442212300_bib1","doi-asserted-by":"publisher","first-page":"931","DOI":"10.1002\/bit.20914","article-title":"Parallel, high-throughput purification of recombinant antibodies for in vivo cell assays","volume":"94","author":"Bannister","year":"2006","journal-title":"Biotechnol. Bioeng."},{"key":"2023072811442212300_bib2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1004952","article-title":"Filopodial-Tension Model of Convergent-Extension of Tissues","volume":"12","author":"Belmonte","year":"2016","journal-title":"PLOS Comput. Biol."},{"key":"2023072811442212300_bib3","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1016\/j.tcb.2016.11.001","article-title":"Endocytosis, Metastasis and Beyond: Multiple Facets of SNX9","volume":"27","author":"Bendris","year":"2017","journal-title":"Trends Cell Biol."},{"key":"2023072811442212300_bib4","doi-asserted-by":"publisher","first-page":"2804","DOI":"10.1242\/jcs.188045","article-title":"Sorting nexin 9 negatively regulates invadopodia formation and function in cancer cells","volume":"129","author":"Bendris","year":"2016","journal-title":"J. Cell Sci."},{"key":"2023072811442212300_bib5","doi-asserted-by":"crossref","DOI":"10.1091\/mbc.E16-02-0101","article-title":"SNX9 promotes metastasis by enhancing cancer cell invasion via differential regulation of RhoGTPases","author":"Bendris","year":"2016","journal-title":"Mol. Biol. Cell."},{"key":"2023072811442212300_bib6","doi-asserted-by":"publisher","first-page":"11622","DOI":"10.1074\/jbc.M805940200","article-title":"The Toca-1-N-WASP complex links filopodial formation to endocytosis","volume":"284","author":"Bu","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2023072811442212300_bib7","doi-asserted-by":"publisher","first-page":"822","DOI":"10.1002\/cm.20226","article-title":"Versatile fluorescent probes for actin filaments based on the actin-binding domain of utrophin","volume":"64","author":"Burkel","year":"2007","journal-title":"Cell Motil. Cytoskeleton."},{"key":"2023072811442212300_bib8","doi-asserted-by":"publisher","first-page":"300","DOI":"10.3389\/fmicb.2016.00300","article-title":"Filopodia and Viruses: An Analysis of Membrane Processes in Entry Mechanisms","volume":"7","author":"Chang","year":"2016","journal-title":"Front. Microbiol."},{"key":"2023072811442212300_bib9","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1016\/j.ydbio.2013.07.024","article-title":"Blastocoel-spanning filopodia in cleavage-stage Xenopus laevis: Potential roles in morphogen distribution and detection","volume":"382","author":"Danilchik","year":"2013","journal-title":"Dev. Biol."},{"key":"2023072811442212300_bib10","doi-asserted-by":"publisher","first-page":"132","DOI":"10.1002\/btpr.1809","article-title":"A high-yielding CHO transient system: coexpression of genes encoding EBNA-1 and GS enhances transient protein expression","volume":"30","author":"Daramola","year":"2014","journal-title":"Biotechnol. Prog."},{"key":"2023072811442212300_bib11","doi-asserted-by":"publisher","first-page":"3745","DOI":"10.1083\/jcb.201704061","article-title":"Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature","volume":"216","author":"Daste","year":"2017","journal-title":"J. Cell Biol."},{"key":"2023072811442212300_bib12","doi-asserted-by":"publisher","first-page":"1347","DOI":"10.1038\/ncb1654","article-title":"Filopodia are required for cortical neurite initiation","volume":"9","author":"Dent","year":"2007","journal-title":"Nat. Cell Biol."},{"key":"2023072811442212300_bib13","doi-asserted-by":"publisher","DOI":"10.1101\/525725","article-title":"Constrained actin dynamics emerges from variable compositions of actin regulatory protein complexes","volume-title":"bioRxiv.","author":"Dobramysl","year":"2019"},{"key":"2023072811442212300_bib14","doi-asserted-by":"publisher","first-page":"2137","DOI":"10.1242\/dev.002345","article-title":"Ena\/VASP function in retinal axons is required for terminal arborization but not pathway navigation","volume":"134","author":"Dwivedy","year":"2007","journal-title":"Development."},{"key":"2023072811442212300_bib15","doi-asserted-by":"publisher","first-page":"1656","DOI":"10.1016\/j.biocel.2009.02.012","article-title":"Filopodia: Complex models for simple rods","volume":"41","author":"Faix","year":"2009","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"2023072811442212300_bib16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1007051","article-title":"Chlamydia exploits filopodial capture and a macropinocytosis-like pathway for host cell entry","volume":"14","author":"Ford","year":"2018","journal-title":"PLoS Pathog."},{"key":"2023072811442212300_bib17","article-title":"Filopodia and their links with membrane traffic and cell adhesion","author":"Gallop","year":"2019","journal-title":"Semin. Cell Dev. Biol."},{"key":"2023072811442212300_bib18","doi-asserted-by":"publisher","first-page":"7193","DOI":"10.1073\/pnas.1305286110","article-title":"Phosphoinositides and membrane curvature switch the mode of actin polymerization via selective recruitment of toca-1 and Snx9","volume":"110","author":"Gallop","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072811442212300_bib19","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1042\/BJ20090428","article-title":"Immunocytochemical techniques reveal multiple, distinct cellular pools of PtdIns4P and PtdIns(4,5)P(2)","volume":"422","author":"Hammond","year":"2009","journal-title":"Biochem. J."},{"key":"2023072811442212300_bib20","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.cub.2018.11.053","article-title":"Filopodome Mapping Identifies p130Cas as a Mechanosensitive Regulator of Filopodia Stability","volume":"29","author":"Jacquemet","year":"2019","journal-title":"Curr. Biol."},{"key":"2023072811442212300_bib21","doi-asserted-by":"publisher","first-page":"1827","DOI":"10.1016\/j.bpj.2016.09.022","article-title":"Are Filopodia Privileged Signaling Structures in Migrating Cells?","volume":"111","author":"Johnson","year":"2016","journal-title":"Biophys. J."},{"key":"2023072811442212300_bib22","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1242\/dev.103.1.193","article-title":"Regional expression, pattern and timing of convergence and extension during gastrulation of Xenopus laevis","volume":"103","author":"Keller","year":"1988","journal-title":"Development."},{"key":"2023072811442212300_bib23","doi-asserted-by":"publisher","first-page":"897","DOI":"10.1098\/rstb.2000.0626","article-title":"Mechanisms of convergence and extension by cell intercalation","volume":"355","author":"Keller","year":"2000","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"2023072811442212300_bib24","doi-asserted-by":"publisher","first-page":"1341","DOI":"10.1126\/science.1191710","article-title":"Self-assembly of filopodia-like structures on supported lipid bilayers","volume":"329","author":"Lee","year":"2010","journal-title":"Science."},{"key":"2023072811442212300_bib25","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1093\/protein\/gzn058","article-title":"Modelling the human immune response: performance of a 1011 human antibody repertoire against a broad panel of therapeutically relevant antigens","volume":"22","author":"Lloyd","year":"2009","journal-title":"Protein Eng. Des. Sel."},{"key":"2023072811442212300_bib26","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.02975","article-title":"A role of OCRL in clathrin-coated pit dynamics and uncoating revealed by studies of Lowe syndrome cells","volume":"3","author":"N\u00e1ndez","year":"2014","journal-title":"eLife."},{"key":"2023072811442212300_bib27","doi-asserted-by":"publisher","first-page":"2203","DOI":"10.1016\/j.celrep.2017.02.008","article-title":"Coordinated Movement of Vesicles and Actin Bundles during Nerve Growth Revealed by Superresolution Microscopy","volume":"18","author":"Nozumi","year":"2017","journal-title":"Cell Rep."},{"key":"2023072811442212300_bib28","doi-asserted-by":"publisher","first-page":"181","DOI":"10.1016\/S1380-2933(96)00046-2","article-title":"Generation of a panel of related human scFv antibodies with high affinities for human CEA","volume":"2","author":"Osbourn","year":"1996","journal-title":"Immunotechnology."},{"key":"2023072811442212300_bib29","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/S0378-1119(96)00628-2","article-title":"An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries","volume":"187","author":"Persic","year":"1997","journal-title":"Gene."},{"key":"2023072811442212300_bib30","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: an open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods."},{"key":"2023072811442212300_bib31","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1083\/jcb.201112113","article-title":"The Arp2\/3 complex is required for lamellipodia extension and directional fibroblast cell migration","volume":"197","author":"Suraneni","year":"2012","journal-title":"J. Cell Biol."},{"key":"2023072811442212300_bib32","first-page":"2579","article-title":"Visualizing data using t-SNE","volume":"9","author":"van der Maaten","year":"2008","journal-title":"J. Mach. Learn. Res."},{"key":"2023072811442212300_bib33","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1038\/nbt0396-309","article-title":"Human antibodies with sub-nanomolar affinities isolated from a large non-immunized phage display library","volume":"14","author":"Vaughan","year":"1996","journal-title":"Nat. Biotechnol."},{"key":"2023072811442212300_bib34","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1016\/bs.mcb.2015.01.020","article-title":"Triggering actin polymerization in Xenopus egg extracts from phosphoinositide-containing lipid bilayers","volume":"128","author":"Walrant","year":"2015","journal-title":"Methods Cell Biol."},{"key":"2023072811442212300_bib35","doi-asserted-by":"publisher","first-page":"973","DOI":"10.1016\/j.cell.2011.12.034","article-title":"Arp2\/3 is critical for lamellipodia and response to extracellular matrix cues but is dispensable for chemotaxis","volume":"148","author":"Wu","year":"2012","journal-title":"Cell."},{"key":"2023072811442212300_bib36","doi-asserted-by":"publisher","first-page":"1467","DOI":"10.1016\/j.cub.2014.05.044","article-title":"Deep proteomics of the Xenopus laevis egg using an mRNA-derived reference database","volume":"24","author":"W\u00fchr","year":"2014","journal-title":"Curr. Biol."},{"key":"2023072811442212300_bib37","doi-asserted-by":"publisher","first-page":"402","DOI":"10.4161\/cam.5.5.16971","article-title":"Filopodia initiation: focus on the Arp2\/3 complex and formins","volume":"5","author":"Yang","year":"2011","journal-title":"Cell Adhes. Migr."},{"key":"2023072811442212300_bib38","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1016\/j.devcel.2007.04.014","article-title":"SNX9 couples actin assembly to phosphoinositide signals and is required for membrane remodeling during endocytosis","volume":"13","author":"Yarar","year":"2007","journal-title":"Dev. Cell."},{"key":"2023072811442212300_bib39","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.220814","article-title":"Roles for Ena\/VASP proteins in FMNL3-mediated filopodial assembly","volume":"131","author":"Young","year":"2018","journal-title":"J. Cell Sci."}],"container-title":["Journal of Cell Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/doi\/10.1083\/jcb.201909178\/1831492\/jcb_201909178.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/doi\/10.1083\/jcb.201909178\/1831492\/jcb_201909178.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,28]],"date-time":"2023-07-28T07:47:24Z","timestamp":1690530444000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jcb\/article\/219\/4\/e201909178\/151579\/A-direct-role-for-SNX9-in-the-biogenesis-of"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,25]]},"references-count":39,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2020,4,6]]}},"URL":"https:\/\/doi.org\/10.1083\/jcb.201909178","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/710285","asserted-by":"object"}]},"ISSN":["0021-9525","1540-8140"],"issn-type":[{"value":"0021-9525","type":"print"},{"value":"1540-8140","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,25]]},"article-number":"e201909178"}}