Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis
Entry of dengue virus 2 (DENV-2) into Aedes albopictus mosquito C6/36 cells was analysed using biochemical and molecular inhibitors, together with confocal and electron microscopy observations. Treatment with monodansylcadaverine, chlorpromazine, sucrose and ammonium chloride inhibited DENV-2 virus...
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2008
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00221317_v89_n2_p474_Acosta http://hdl.handle.net/20.500.12110/paper_00221317_v89_n2_p474_Acosta |
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paper:paper_00221317_v89_n2_p474_Acosta2023-06-08T14:46:33Z Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis actin ammonium chloride chlorpromazine clathrin colchicine cytochalasin D dansylcadaverine nystatin sucrose transferrin virus glycoprotein clathrin cytochalasin D virus envelope protein Aedes albopictus animal cell article cell membrane coated pit coated vesicle confocal microscopy controlled study Dengue virus dengue virus 2 electron microscopy endocytosis endosome Flavivirus microtubule mosquito nonhuman pH priority journal protein expression protein localization virus entry virus infectivity virus particle virus replication Aedes animal Cercopithecus drug effect genetics metabolism microfilament physiology transport at the cellular level ultrastructure Vero cell virology Aedes albopictus Dengue virus Hexapoda Actins Aedes Animals Biological Transport Cercopithecus aethiops Clathrin Cytochalasin D Dengue Virus Endocytosis Endosomes Microfilaments Microscopy, Confocal Microtubules Transferrin Vero Cells Viral Envelope Proteins Virus Internalization Entry of dengue virus 2 (DENV-2) into Aedes albopictus mosquito C6/36 cells was analysed using biochemical and molecular inhibitors, together with confocal and electron microscopy observations. Treatment with monodansylcadaverine, chlorpromazine, sucrose and ammonium chloride inhibited DENV-2 virus yield and protein expression, whereas nystatin, a blocker of caveolae-mediated endocytosis, did not have any effect. Using confocal microscopy, co-localization of DENV-2 E glycoprotein and the marker protein transferrin was observed at the periphery of the cytoplasm. To support the requirement of clathrin function for DENV-2 entry, overexpression of a dominant-negative mutant of Eps15 in C6/36 cells was shown to impair virus entry. The disruption of actin microfilaments by cytochalasin D also significantly affected DENV-2 replication. In contrast, microtubule disruption by colchicine treatment did not impair DENV-2 infectivity, suggesting that DENV-2 does not require transport from early to late endosomes for successful infection of mosquito cells. Furthermore, using transmission electron microscopy, DENV-2 particles of approximately 44-52 nm were found attached within electron-dense invaginations of the plasma membrane and in coated vesicles that resembled those of clathrin-coated pits and vesicles, respectively. Together, these results demonstrate for the first time that DENV-2 enters insect cells by receptor-mediated, clathrin-dependent endocytosis, requiring traffic through an acidic pH compartment for subsequent uncoating and completion of a productive infection. © 2008 SGM. 2008 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00221317_v89_n2_p474_Acosta http://hdl.handle.net/20.500.12110/paper_00221317_v89_n2_p474_Acosta |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
actin ammonium chloride chlorpromazine clathrin colchicine cytochalasin D dansylcadaverine nystatin sucrose transferrin virus glycoprotein clathrin cytochalasin D virus envelope protein Aedes albopictus animal cell article cell membrane coated pit coated vesicle confocal microscopy controlled study Dengue virus dengue virus 2 electron microscopy endocytosis endosome Flavivirus microtubule mosquito nonhuman pH priority journal protein expression protein localization virus entry virus infectivity virus particle virus replication Aedes animal Cercopithecus drug effect genetics metabolism microfilament physiology transport at the cellular level ultrastructure Vero cell virology Aedes albopictus Dengue virus Hexapoda Actins Aedes Animals Biological Transport Cercopithecus aethiops Clathrin Cytochalasin D Dengue Virus Endocytosis Endosomes Microfilaments Microscopy, Confocal Microtubules Transferrin Vero Cells Viral Envelope Proteins Virus Internalization |
| spellingShingle |
actin ammonium chloride chlorpromazine clathrin colchicine cytochalasin D dansylcadaverine nystatin sucrose transferrin virus glycoprotein clathrin cytochalasin D virus envelope protein Aedes albopictus animal cell article cell membrane coated pit coated vesicle confocal microscopy controlled study Dengue virus dengue virus 2 electron microscopy endocytosis endosome Flavivirus microtubule mosquito nonhuman pH priority journal protein expression protein localization virus entry virus infectivity virus particle virus replication Aedes animal Cercopithecus drug effect genetics metabolism microfilament physiology transport at the cellular level ultrastructure Vero cell virology Aedes albopictus Dengue virus Hexapoda Actins Aedes Animals Biological Transport Cercopithecus aethiops Clathrin Cytochalasin D Dengue Virus Endocytosis Endosomes Microfilaments Microscopy, Confocal Microtubules Transferrin Vero Cells Viral Envelope Proteins Virus Internalization Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| topic_facet |
actin ammonium chloride chlorpromazine clathrin colchicine cytochalasin D dansylcadaverine nystatin sucrose transferrin virus glycoprotein clathrin cytochalasin D virus envelope protein Aedes albopictus animal cell article cell membrane coated pit coated vesicle confocal microscopy controlled study Dengue virus dengue virus 2 electron microscopy endocytosis endosome Flavivirus microtubule mosquito nonhuman pH priority journal protein expression protein localization virus entry virus infectivity virus particle virus replication Aedes animal Cercopithecus drug effect genetics metabolism microfilament physiology transport at the cellular level ultrastructure Vero cell virology Aedes albopictus Dengue virus Hexapoda Actins Aedes Animals Biological Transport Cercopithecus aethiops Clathrin Cytochalasin D Dengue Virus Endocytosis Endosomes Microfilaments Microscopy, Confocal Microtubules Transferrin Vero Cells Viral Envelope Proteins Virus Internalization |
| description |
Entry of dengue virus 2 (DENV-2) into Aedes albopictus mosquito C6/36 cells was analysed using biochemical and molecular inhibitors, together with confocal and electron microscopy observations. Treatment with monodansylcadaverine, chlorpromazine, sucrose and ammonium chloride inhibited DENV-2 virus yield and protein expression, whereas nystatin, a blocker of caveolae-mediated endocytosis, did not have any effect. Using confocal microscopy, co-localization of DENV-2 E glycoprotein and the marker protein transferrin was observed at the periphery of the cytoplasm. To support the requirement of clathrin function for DENV-2 entry, overexpression of a dominant-negative mutant of Eps15 in C6/36 cells was shown to impair virus entry. The disruption of actin microfilaments by cytochalasin D also significantly affected DENV-2 replication. In contrast, microtubule disruption by colchicine treatment did not impair DENV-2 infectivity, suggesting that DENV-2 does not require transport from early to late endosomes for successful infection of mosquito cells. Furthermore, using transmission electron microscopy, DENV-2 particles of approximately 44-52 nm were found attached within electron-dense invaginations of the plasma membrane and in coated vesicles that resembled those of clathrin-coated pits and vesicles, respectively. Together, these results demonstrate for the first time that DENV-2 enters insect cells by receptor-mediated, clathrin-dependent endocytosis, requiring traffic through an acidic pH compartment for subsequent uncoating and completion of a productive infection. © 2008 SGM. |
| title |
Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| title_short |
Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| title_full |
Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| title_fullStr |
Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| title_full_unstemmed |
Functional entry of dengue virus into Aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| title_sort |
functional entry of dengue virus into aedes albopictus mosquito cells is dependent on clathrin-mediated endocytosis |
| publishDate |
2008 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00221317_v89_n2_p474_Acosta http://hdl.handle.net/20.500.12110/paper_00221317_v89_n2_p474_Acosta |
| _version_ |
1768546382166097920 |