Echovirus 1 internalization negatively regulates epidermal growth factor receptor downregulation.
We have demonstrated previously that the human picornavirus Echovirus 1 (EV1) triggers an infectious internalization pathway that follows closely, but seems to stay separate, from the epidermal growth factor receptor (EGFR) pathway triggered by epidermal growth factor (EGF). Here, we confirmed by using live and confocal microscopy that EGFR and EV1 vesicles are following intimately each other but are distinct entities with different degradation kinetics. We show here that despite being sorted to different pathways and located in distinct endosomes, EV1 inhibits EGFR downregulation. Simultaneous treatment with EV1 and EGF led to an accumulation of EGFR in cytoplasmic endosomes, which was evident already 15 min p.i. and more pronounced after 2 hr p.i. EV1 treatment led to reduced downregulation, which was proven by increased total cellular amount of EGFR. Confocal microscopy studies revealed that EGFR accumulated in large endosomes, presumably macropinosomes, which were not positive for markers of the early, recycling, or late endosomes/lysosomes. Interestingly, EV1 did not have a similar blocking effect on bulk endosomal trafficking or transferrin recycling along the clathrin pathway suggesting that EV1 did not have a general effect on cellular trafficking pathways. Importantly, EGF treatment increased EV1 infection and increased cell viability during infection. Simultaneous EV1 and EGF treatment seemed to moderately enhance phosphorylation of protein kinase C α. Furthermore, similar phenotype of EGFR trafficking could be produced by phorbol 12-myristate 13-acetate treatment, further suggesting that activated protein kinase C α could be contributing to EGFR phenotype. These results altogether demonstrate that EV1 specifically affects EGFR trafficking, leading to EGFR downregulation, which is beneficial to EV1 infection.
Department of Biological and Environmental Science/NanoScience Center, University of Jyväskylä, Jyväskylä, Finland.
Moona
M
Huttunen
Department of Biological and Environmental Science/NanoScience Center, University of Jyväskylä, Jyväskylä, Finland.
Paula
P
Turkki
Department of Biological and Environmental Science/NanoScience Center, University of Jyväskylä, Jyväskylä, Finland.
Anita
A
Mäki
Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland.
Lassi
L
Paavolainen
Institute of Biosciences and Medical Technology, Tampere University of Technology, Pori-University of Tampere, Tampere, Finland.
Pekka
P
Ruusuvuori
Department of Biological and Environmental Science/NanoScience Center, University of Jyväskylä, Jyväskylä, Finland.
Varpu
V
Marjomäki
D002460:Cell Line; D015536:Down-Regulation; D011992:Endosomes / Q000378:metabolism; D029822:Enterovirus B, Human / Q000502:physiology*; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology; D066246:ErbB Receptors / Q000096:biosynthesis*; D054884:Host-Pathogen Interactions; D006801:Humans; D018613:Microscopy, Confocal; D053586:Virus Internalization
D066246:ErbB Receptors