MyosinVb tail inhibits transport of Marburg virus glycoprotein GP to VP40-enriched sites at the plasma membrane.

Marburg virus (MARV) is the causative agent of severe fever with case fatality rates between 25 and 90 %. The glycoprotein GP is the only surface protein of MARV responsible for receptor recognition and fusion. Therefore, proper intracellular transport of GP to the plasma membrane and incorporation into virus particles is essential for the viral infection cycle. However, neither the exact post-Golgi trafficking route nor the host factors are known that support the transport of GP to the cell surface. Using quantitative confocal microscopy and correlative light and electron microscopy (CLEM), we show here that GP colocalized in both transiently transfected and MARV-infected cells with a dominant negative (DN) tail mutant of myosin Vb (MyoVbT), which inhibits trafficking through recycling endosomes. Overexpression of MyoVbT resulted in an aberrant distribution of GP that accumulated in or near perinuclear MyoVbT-containing structures. Simultaneously, we observed significantly reduced GP levels at the plasma membrane and especially at the viral budding sites characterized by clusters of the viral matrix protein VP40. Further, incorporation of GP into VP40-induced filamentous virus-like particles was impaired by MyoVbT. Overall, our results show that intracellular transport of MARV GP is disrupted by a DN mutant of the recycling endosome-associated motor protein MyoVb. These results might indicate a possible role for the endosomal recycling system in MARV GP trafficking to VP40-enriched budding-sites at the plasma membrane.