title: Role of endothelial cell septin 7 in the endocytosis of Candida albicans.
abstract: Candida albicans invades endothelial cells by binding to N-cadherin and other cell surface receptors. This binding induces rearrangement of endothelial cell actin microfilaments, which results in the formation of pseudopods that surround the organism and pull it into the endothelial cell. Here, we investigated the role of endothelial cell septin 7 (SEPT7) in the endocytosis of C. albicans hyphae. Using confocal microscopy, we determined that SEPT7 accumulated with N-cadherin and actin microfilaments around C. albicans as it was endocytosed by endothelial cells. Affinity purification studies indicated that a complex containing N-cadherin and SEPT7 was recruited by C. albicans and that formation of this complex around C. albicans was mediated by the fungal Als3 and Ssa1 invasins. Knockdown of N-cadherin by small interfering RNA (siRNA) reduced recruitment of SEPT7 to C. albicans, suggesting that N-cadherin functions as a link between SEPT7 and the fungus. Also, depolymerization of actin microfilaments with cytochalasin D decreased the association between SEPT7 and N-cadherin and inhibited recruitment of both SEPT7 and N-cadherin to C. albicans, indicating the necessity of an intact cytoskeleton in the functional interaction between SEPT7 and N-cadherin. Importantly, knockdown of SEPT7 decreased accumulation of N-cadherin around C. albicans in intact endothelial cells and reduced binding of N-cadherin to this organism, as revealed by the affinity purification assay. Furthermore, SEPT7 knockdown significantly inhibited the endocytosis of C. albicans. Therefore, in response to C. albicans infection, SEPT7 forms a complex with endothelial cell N-cadherin, is required for normal accumulation of N-cadherin around C. albicans hyphae, and is necessary for maximal endocytosis of the organism.
  
  
  OBJECTIVE
  During hematogenously disseminated infection, Candida albicans invades the endothelial cell lining of the blood vessels to invade the deep tissues. C. albicans can invade endothelial cells by inducing its own endocytosis, which is triggered when the C. albicans Als3 and Ssa1 invasins bind to N-cadherin on the endothelial cell surface. How this binding induces endocytosis is incompletely understood. Septins are intracellular GTP-binding proteins that influence the function and localization of cell surface proteins. We found that C. albicans Als3 and Ssa1 bind to a complex containing N-cadherin and septin 7, which in turn interacts with endothelial cell microfilaments, thereby inducing endocytosis of the organism. The key role of septin 7 in governing receptor-mediated endocytosis is likely relevant to host cell invasion by other microbial pathogens, in addition to C. albicans.
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, California, USA.
authors.forename: Quynh T
authors.initials: QT
authors.lastname: Phan
authors.forename: David K
authors.initials: DK
authors.lastname: Eng
authors.forename: Serge
authors.initials: S
authors.lastname: Mostowy
authors.forename: Hyunsook
authors.initials: H
authors.lastname: Park
authors.forename: Pascale
authors.initials: P
authors.lastname: Cossart
authors.forename: Scott G
authors.initials: SG
authors.lastname: Filler
mesh_terms: D000199:Actins / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism*; D002176:Candida albicans / Q000502:physiology*; D002448:Cell Adhesion; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D004705:Endocytosis; D042783:Endothelial Cells / Q000382:microbiology* / Q000502:physiology*; D005656:Fungal Proteins / Q000378:metabolism; D006801:Humans; D025301:Hyphae / Q000502:physiology; D011485:Protein Binding; D055503:Protein Multimerization; D058112:Septins / Q000378:metabolism*
chemical_list: C113620:ALS3 protein, Candida albicans; D000199:Actins; D015703:Antigens, CD; C050361:CDH2 protein, human; D015820:Cadherins; D018797:Cell Cycle Proteins; D005656:Fungal Proteins; C113158:SEPTIN7 protein, human; D058112:Septins