Viral liposomes released from insect cells infected with recombinant baculovirus expressing the matrix protein of vesicular stomatitis virus.
The matrix (M) protein of vesicular stomatitis virus (VSV) has been found to promote assembly and budding of virions as well as down-regulating of VSV transcription. Large quantities of M protein can be produced in insect cells infected with recombinant baculovirus expressing the VSV M gene under control of the polyhedrin promoter. Analysis by pulse-chase experiments and density gradient centrifugation revealed that the [35S]methionine-labeled M protein synthesized in insect cells is released into the extracellular medium in association with lipid vesicles (liposomes). Electron microscopy and immunogold labeling showed that M protein expressed in insect cells induced the formation on plasma membrane of vesicles containing M protein, which are released from the cell surface in the form of liposomes. The baculovirus vector itself or recombinants expressing VSV glycoprotein (G) or nucleocapsid (N) protein did not produce the formation of vesicles in infected cells. The baculovirus-expressed M protein retains biological activity as demonstrated by its capacity to inhibit transcription when reconstituted with VSV nucleocapsids in vitro. These data suggest that M protein has the capacity to associate with the plasma membrane of infected cells and, in so doing, causes evagination of the membrane to form a vesicle which is released from the cell. This observation leads to the postulate, which requires further proof, that the VSV M protein can induce the formation and budding of liposomes from the cell membrane surface.
Li
Y
Y
Department of Zoology, Faculty of Science, University of Western Ontario, London, Canada.
Luo
L
L
Schubert
M
M
Wagner
R R
RR
Kang
C Y
CY
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular; D066298:In Vitro Techniques; D008081:Liposomes; D008854:Microscopy, Electron; D008970:Molecular Weight; D009036:Moths; D011994:Recombinant Proteins / Q000737:chemistry; D014721:Vesicular stomatitis Indiana virus / Q000737:chemistry*; D014763:Viral Matrix Proteins / Q000096:biosynthesis*
D008081:Liposomes; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins