Permeability changes of integrin-containing multivesicular structures triggered by picornavirus entry.

• Main Authors: Pan Soonsawad, แพน สุ่นสวัสดิ์, Wattana Weerachatyanukul, วัฒนา วีรชาติยานุกูล, Paavolainen, Lassi, Upla, Paula, Rintanen, Nina, Espinoza, Juan, McNerney, Gregory, Marjomäki, Varpu, Cheng, R. Holland
• Other Authors: Mahidol University. Faculty of Dentistry. Department of Anatomy
• Format: Article
• Language: English
• Published: 2015
• Subjects: Cell membranes; Permeability; Endosomes; Antibodies; Integrins; Membrane structures; Vesicles; Virus uncoating; Open Access article
• Online Access: https://repository.li.mahidol.ac.th/handle/123456789/937
• Institution: Mahidol University

Cellular uptake of clustered α2β1-integrin induces the formation of membrane compartments that subsequently mature into a multivesicular body (MVB). Enhanced internalization mediated by clustered integrins was observed upon infection by the picornavirus echovirus 1 (EVI). We elucidated the structural features of virus-induced MVBs (vMVBs) in comparison to antibody-induced control MVBs (mock infection) by means of high-pressure cryo fixation of cells followed by immuno electron tomography during early entry of the virus. Three-dimensional tomograms revealed a marked increase in the size and complexity of these vMVBs and the intraluminal vesicles (ILVs) at 2 and 3.5 hours post infection (p.i.), in contrast to the control MVBs without virus. Breakages in the membranes of vMVBs were detected from tomograms after 2 and especially after 3.5 h suggesting that these breakages could facilitate the genome release to the cytoplasm. The in situ neutral-red labeling of viral genome showed that virus uncoating starts as early as 30 min p.i., while an increase of permeability was detected in the vMVBs between 1 and 3 hours p.i., based on a confocal microscopy assay. Altogether, the data show marked morphological changes in size and permeability of the endosomes in the infectious entry pathway of this non-enveloped enterovirus and suggest that the formed breakages facilitate the transfer of the genome to the cytoplasm for replication.