ERK MAP kinase-activated Arf6 trafficking directs coxsackievirus type B3 into an unproductive compartment during virus host-cell entry.
Clathrin- and caveolae-mediated endocytosis have been implicated in the productive entry of many viruses into host cells. ADP-ribosylation factor 6 (Arf6)-dependent endocytosis is another endocytosis pathway that traffics from the cell surface and it is the only Arf that traffics at the plasma membrane. However, little is known about Arf6-dependent trafficking during virus entry. This study showed that coxsackievirus type B3 (CVB3) associated with decay-accelerating factor in non-polarized HeLa cells can be redirected into non-productive compartments by Arf6-dependent internalization, thus restricting infection. Overexpression of wild-type (WT) and constitutively active (CA) Arf6 in HeLa cells resulted in a 2.3- and 3.6-fold decrease in infection, respectively. A dominant-negative inhibitor of Arf6 recovered restriction of infection by WT-Arf6 and CA-Arf6. RNA interference of endogenous Arf6 resulted in a 3.3-fold increase in CVB3 titre in HeLa cells. It was shown that coxsackie-adenovirus receptor (CAR) ligation by virus or CAR-specific antibody could activate extracellular signal-regulated kinase (ERK) of the mitogen-activated protein kinase family and lead to Arf6-mediated viral restriction. In the absence of ERK activation, CVB3 internalization into early endosomes was inhibited and subsequent infection was reduced, but Arf6-mediated restriction was also abolished. In conclusion, receptor-mediated signalling enhances CVB3 entry whilst also activating non-productive pathways of virus entry; thus, virus infection is an equilibrium of productive and non-productive pathways of entry.
Marchant
David
D
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Sall
Alhousseynou
A
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Si
Xiaoning
X
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Abraham
Thomas
T
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Wu
Winnie
W
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Luo
Zongshu
Z
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Petersen
Tamar
T
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
Hegele
Richard G
RG
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
McManus
Bruce M
BM
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, Departments of Pathology and Laboratory Medicine, The University of British Columbia, Room 166 Burrard Building, St Paul's Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, Canada.
D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004705:Endocytosis; D011992:Endosomes / Q000821:virology; D029822:Enterovirus B, Human / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D000089982:ADP-Ribosylation Factor 6; C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011991:Receptors, Virus; D048049:Extracellular Signal-Regulated MAP Kinases; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human