Immunolocalization studies of vimentin and ACE2 on the surface of cells exposed to SARS-CoV-2 Spike proteins

Autor: Dolores Pérez-Sala, Irene Lois-Bermejo, Silvia González-Sanz, Álvaro Viedma-Poyatos, María A. Pajares, Vasiliky S. Lalioti, Patricia González-Jiménez, Andrea Merino
Přispěvatelé: Consejo Superior de Investigaciones Científicas (España), Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), Pajares, María A., Pérez-Sala, Dolores, Pajares, María A. [0000-0002-4714-9051], Pérez-Sala, Dolores [0000-0003-0600-665X]
Rok vydání: 2021
Předmět:
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
Digital.CSIC: Repositorio Institucional del CSIC
Consejo Superior de Investigaciones Científicas (CSIC)
DOI: 10.1101/2021.05.04.442648
Popis: The Spike protein from SARS-CoV-2 mediates docking of the virus onto cells and contributes to viral invasion. Several cellular receptors are involved in SARS-CoV-2 Spike docking at the cell surface, including ACE2 and neuropilin. The intermediate filament protein vimentin has been reported to be present at the surface of certain cells and act as a co-receptor for several viruses; furthermore, its potential involvement in interactions with Spike proteins has been proposed. Here we have explored the binding of Spike protein constructs to several cell types using low-temperature immunofluorescence approaches in live cells, to minimize internalization. Incubation of cells with tagged Spike S or Spike S1 subunit led to discrete dotted patterns at the cell surface, which showed scarce colocalization with a lipid raft marker, but consistent coincidence with ACE2. Under our conditions, vimentin immunoreactivity appeared as spots or patches unevenly distributed at the surface of diverse cell types. Remarkably, several observations including potential antibody internalization and adherence to cells of vimentin-positive structures present in the extracellular medium exposed the complexity of vimentin cell surface immunoreactivity, which requires careful assessment. Notably, overall colocalization of Spike and vimentin signals markedly varied with the cell type and the immunodetection sequence. In turn, vimentin-positive spots moderately colocalized with ACE2; however, a particular enrichment was detected at elongated structures positive for acetylated tubulin, consistent with primary cilia, which also showed Spike binding. Thus, these results suggest that vimentin-ACE2 interaction could occur at selective locations near the cell surface, including ciliated structures, which can act as platforms for SARS-CoV-2 docking.
This work was supported by grants CSIC PTI Global Health (PIE 202020E223/CSIC-COV19-100), RTI2018-097624-B-I00 from Micinn (Agencia Estatal de Investigación), Spain and ERDF. Á.V.P. and P.G.J. are the recipients of predoctoral contracts BES-2016-076965 and PRE2019-088194, respectively, from Micinn, Spain.
Databáze: OpenAIRE
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