A Myosin Vb Splice Variant Regulates Coronavirus M Protein Trafficking in Polarized Epithelial Cells

James R. Goldenring, Lynne A. Lapierre, Joseph T. Roland, Elizabeth Manning, Cathy Caldwell, Honor L. Glenn, Pierre Olivier Vidalain, Frederic Tangy, Brenda G. Hogue, C. A.M. de Haan

Research output: Contribution to journalArticlepeer-review

Abstract

The membrane (M) glycoprotein of coronaviruses (CoVs) serves as the nidus for virion assembly. Using a yeast two-hybrid screen, we identified the interaction of the cytosolic tail of Murine Hepatitis Virus (MHV-CoV) M protein with Myosin Vb (MYO5B) (Figure 1A). MHV-CoV M interacts specifically with the alternative splice variant of cellular MYO5B including Exon D (MYO5B+D), that also mediates interaction with cellular Rab10. When co-expressed in human lung epithelial A549 and canine kidney epithelial MDCK cells, MYO5B+D co-localized with MHV-CoV M protein, as well as with M proteins from porcine epidemic diarrhea Virus (PEDV-CoV), Middle East Respiratory Syndrome (MERS-CoV) and Severe Acute Respiratory Syndrome 2 (SARS-CoV-2) (Figure 2). M-GFP chimeric proteins co-expressed with mCherry-MYO5B+D also co-localized with endogenous Rab10 and Rab11a (Figure 1B,C). We identified point mutations in MHV-CoV M that blocked the interaction with MYO5B+D in yeast 2-hybrid assays (Figure 1B). One of these point mutations (E121K) was previously shown to block MHV-CoV virion assembly, and it blocked interaction with MYO5B+D. The E to K mutation at homologous positions in PEDV-CoV, MERS-CoV and SARS-CoV-2 M proteins also blocked colocalization with MYO5B+D. Knockdown of Rab10 blocked the co-localization of the M proteins with MYO5B+D (Figure 2). Re-expression of Cerulein-Rab10 in Rab10 KD cells re-established colocalization between M proteins and MYO5B+D (Figure 2). Our results suggest that the interaction of CoV M proteins with MYO5B+D may play a role in regulating their trafficking through Rab10-containing membrane systems in epithelial cells.

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Fingerprint

Dive into the research topics of 'A Myosin Vb Splice Variant Regulates Coronavirus M Protein Trafficking in Polarized Epithelial Cells'. Together they form a unique fingerprint.

Cite this