Abstract
The matrix (M) protein of vesicular stomatitis virus (VSV) is a major structural component of the virion which is generally believed to bridge between the membrane envelope and the ribonucleocapsid (RNP) core. To investigate the interaction of M protein with cellular membranes in the absence of other VSV proteins, we examined its distribution by subcellular fractionation after expression in HeLa cells. Approximately 90% of M protein, expressed without other viral proteins, was soluble, whereas the remaining 10% was tightly associated with membranes. A similar distribution in VSV-infected cells has been observed previously. Conditions known to release peripherally associated membrane proteins did not detach M protein from isolated membranes. Membrane-associated M protein was soluble in the detergent Triton X-114, whereas soluble M protein was not, suggesting a chemical or conformational difference between the two forms. Membranes containing associated M protein were able to bind RNP cores, whereas membranes lacking M protein were not. We suggest that this membrane-bound M fraction constitutes a functional subset of M protein molecules required for the attachment of RNP cores to membranes during normal virus budding.
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- Atkinson P. H. Gycoprotein and protein precursors to plasma membranes in vesicular stomatitis virus infected HeLa cells. J Supramol Struct. 1978;8(1):89–109. doi: 10.1002/jss.400080108. [DOI] [PubMed] [Google Scholar]
- Atkinson P. H., Moyer S. A., Summers D. F. Assembly of vesicular stomatitis virus glycoprotein and matrix protein into HeLa cell plasma membranes. J Mol Biol. 1976 Apr 15;102(3):613–631. doi: 10.1016/0022-2836(76)90338-7. [DOI] [PubMed] [Google Scholar]
- Bergmann J. E., Fusco P. J. The M protein of vesicular stomatitis virus associates specifically with the basolateral membranes of polarized epithelial cells independently of the G protein. J Cell Biol. 1988 Nov;107(5):1707–1715. doi: 10.1083/jcb.107.5.1707. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
- Bucher D. J., Kharitonenkov I. G., Zakomirdin J. A., Grigoriev V. B., Klimenko S. M., Davis J. F. Incorporation of influenza virus M-protein into liposomes. J Virol. 1980 Nov;36(2):586–590. doi: 10.1128/jvi.36.2.586-590.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Caldwell S. E., Lyles D. S. Dissociation of newly synthesized Sendai viral proteins from the cytoplasmic surface of isolated plasma membranes of infected cells. J Virol. 1986 Feb;57(2):678–683. doi: 10.1128/jvi.57.2.678-683.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Capone J., Ghosh H. P. Association of the nucleocapsid protein N of vesicular stomatitis virus with phospholipid vesicles containing the matrix protein M. Can J Biochem Cell Biol. 1984 Nov;62(11):1174–1180. doi: 10.1139/o84-151. [DOI] [PubMed] [Google Scholar]
- Carroll A. R., Wagner R. R. Role of the membrane (M) protein in endogenous inhibition of in vitro transcription by vesicular stomatitis virus. J Virol. 1979 Jan;29(1):134–142. doi: 10.1128/jvi.29.1.134-142.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen S. S., Ariel N., Huang A. S. Membrane anchors of vesicular stomatitis virus: characterization and incorporation into virions. J Virol. 1988 Aug;62(8):2552–2556. doi: 10.1128/jvi.62.8.2552-2556.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clinton G. M., Burge B. W., Huang A. S. Effects of phosphorylation and pH on the association of NS protein with vesicular stomatitis virus cores. J Virol. 1978 Aug;27(2):340–346. doi: 10.1128/jvi.27.2.340-346.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cohen G. H., Summers D. F. In vitro association of vesicular stomatitis virus proteins with purified HeLa and erythrocyte plasma membranes. Virology. 1974 Feb;57(2):566–569. doi: 10.1016/0042-6822(74)90195-0. [DOI] [PubMed] [Google Scholar]
- David A. E. Assembly of the vesicular stomatitis virus envelope: incorporation of viral polypeptides into the host plasma membrane. J Mol Biol. 1973 May 5;76(1):135–148. doi: 10.1016/0022-2836(73)90085-5. [DOI] [PubMed] [Google Scholar]
- De B. P., Thornton G. B., Luk D., Banerjee A. K. Purified matrix protein of vesicular stomatitis virus blocks viral transcription in vitro. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7137–7141. doi: 10.1073/pnas.79.23.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dubovi E. J., Wagner R. R. Spatial relationships of the proteins of vesicular stomatitis virus: induction of reversible oligomers by cleavable protein cross-linkers and oxidation. J Virol. 1977 May;22(2):500–509. doi: 10.1128/jvi.22.2.500-509.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Faaberg K. S., Peeples M. E. Association of soluble matrix protein of Newcastle disease virus with liposomes is independent of ionic conditions. Virology. 1988 Sep;166(1):123–132. doi: 10.1016/0042-6822(88)90153-5. [DOI] [PubMed] [Google Scholar]
- Fuerst T. R., Niles E. G., Studier F. W., Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122–8126. doi: 10.1073/pnas.83.21.8122. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fujiki Y., Hubbard A. L., Fowler S., Lazarow P. B. Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biol. 1982 Apr;93(1):97–102. doi: 10.1083/jcb.93.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gregoriades A. Interaction of influenza M protein with viral lipid and phosphatidylcholine vesicles. J Virol. 1980 Nov;36(2):470–479. doi: 10.1128/jvi.36.2.470-479.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hua Q. X., Kochoyan M., Weiss M. A. Structure and dynamics of des-pentapeptide-insulin in solution: the molten-globule hypothesis. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2379–2383. doi: 10.1073/pnas.89.6.2379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jacobs B. L., Penhoet E. E. Assembly of vesicular stomatitis virus: distribution of the glycoprotein on the surface of infected cells. J Virol. 1982 Dec;44(3):1047–1055. doi: 10.1128/jvi.44.3.1047-1055.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Khan S. R., Lazzarini R. A. The relationship between autointerference and the replication of defective interfering particle. Virology. 1977 Mar;77(1):189–201. doi: 10.1016/0042-6822(77)90417-2. [DOI] [PubMed] [Google Scholar]
- Knipe D. M., Baltimore D., Lodish H. F. Maturation of viral proteins in cells infected with temperature-sensitive mutants of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1149–1158. doi: 10.1128/jvi.21.3.1149-1158.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knipe D. M., Baltimore D., Lodish H. F. Separate pathways of maturation of the major structural proteins of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1128–1139. doi: 10.1128/jvi.21.3.1128-1139.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lenard J., Vanderoef R. Localization of the membrane-associated region of vesicular stomatitis virus M protein at the N terminus, using the hydrophobic, photoreactive probe 125I-TID. J Virol. 1990 Jul;64(7):3486–3491. doi: 10.1128/jvi.64.7.3486-3491.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lyles D. S., McKenzie M., Parce J. W. Subunit interactions of vesicular stomatitis virus envelope glycoprotein stabilized by binding to viral matrix protein. J Virol. 1992 Jan;66(1):349–358. doi: 10.1128/jvi.66.1.349-358.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mancarella D. A., Lenard J. Interactions of wild-type and mutant M protein of vesicular stomatitis virus with viral nucleocapsid and envelope in intact virions. Evidence from [125I]iodonaphthyl azide labeling and specific cross-linking. Biochemistry. 1981 Nov 24;20(24):6872–6877. doi: 10.1021/bi00527a020. [DOI] [PubMed] [Google Scholar]
- McCreedy B. J., Jr, Lyles D. S. Distribution of M protein and nucleocapsid protein of vesicular stomatitis virus in infected cell plasma membranes. Virus Res. 1989 Nov;14(3):189–205. doi: 10.1016/0168-1702(89)90001-4. [DOI] [PubMed] [Google Scholar]
- McCreedy B. J., Jr, McKinnon K. P., Lyles D. S. Solubility of vesicular stomatitis virus M protein in the cytosol of infected cells or isolated from virions. J Virol. 1990 Feb;64(2):902–906. doi: 10.1128/jvi.64.2.902-906.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Metsikkö K., Simons K. The budding mechanism of spikeless vesicular stomatitis virus particles. EMBO J. 1986 Aug;5(8):1913–1920. doi: 10.1002/j.1460-2075.1986.tb04444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morrison T. G., McQuain C. O. Assembly of viral membranes: nature of the association of vesicular stomatitis virus proteins to membranes. J Virol. 1978 Apr;26(1):115–125. doi: 10.1128/jvi.26.1.115-125.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mudd J. A., Swanson R. E. In situ cross-linking of vesicular stomatitis virus proteins with reversible agents. Virology. 1978 Jul 15;88(2):263–280. doi: 10.1016/0042-6822(78)90284-2. [DOI] [PubMed] [Google Scholar]
- Newcomb W. W., Brown J. C. Role of the vesicular stomatitis virus matrix protein in maintaining the viral nucleocapsid in the condensed form found in native virions. J Virol. 1981 Jul;39(1):295–299. doi: 10.1128/jvi.39.1.295-299.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Newcomb W. W., Tobin G. J., McGowan J. J., Brown J. C. In vitro reassembly of vesicular stomatitis virus skeletons. J Virol. 1982 Mar;41(3):1055–1062. doi: 10.1128/jvi.41.3.1055-1062.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Odenwald W. F., Arnheiter H., Dubois-Dalcq M., Lazzarini R. A. Stereo images of vesicular stomatitis virus assembly. J Virol. 1986 Mar;57(3):922–932. doi: 10.1128/jvi.57.3.922-932.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ogden J. R., Pal R., Wagner R. R. Mapping regions of the matrix protein of vesicular stomatitis virus which bind to ribonucleocapsids, liposomes, and monoclonal antibodies. J Virol. 1986 Jun;58(3):860–868. doi: 10.1128/jvi.58.3.860-868.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ono K., Dubois-Dalcq M. E., Schubert M., Lazzarini R. A. A mutated membrane protein of vesicular stomatitis virus has an abnormal distribution within the infected cell and causes defective budding. J Virol. 1987 May;61(5):1332–1341. doi: 10.1128/jvi.61.5.1332-1341.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reidler J. A., Keller P. M., Elson E. L., Lenard J. A fluorescence photobleaching study of vesicular stomatitis virus infected BHK cells. Modulation of G protein mobility by M protein. Biochemistry. 1981 Mar 3;20(5):1345–1349. doi: 10.1021/bi00508a047. [DOI] [PubMed] [Google Scholar]
- Rose J. K., Bergmann J. E. Altered cytoplasmic domains affect intracellular transport of the vesicular stomatitis virus glycoprotein. Cell. 1983 Sep;34(2):513–524. doi: 10.1016/0092-8674(83)90384-7. [DOI] [PubMed] [Google Scholar]
- Rose J. K., Gallione C. J. Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus G and M proteins determined from cDNA clones containing the complete coding regions. J Virol. 1981 Aug;39(2):519–528. doi: 10.1128/jvi.39.2.519-528.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sprague J., Condra J. H., Arnheiter H., Lazzarini R. A. Expression of a recombinant DNA gene coding for the vesicular stomatitis virus nucleocapsid protein. J Virol. 1983 Feb;45(2):773–781. doi: 10.1128/jvi.45.2.773-781.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stigter D., Alonso D. O., Dill K. A. Protein stability: electrostatics and compact denatured states. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4176–4180. doi: 10.1073/pnas.88.10.4176. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas D., Newcomb W. W., Brown J. C., Wall J. S., Hainfeld J. F., Trus B. L., Steven A. C. Mass and molecular composition of vesicular stomatitis virus: a scanning transmission electron microscopy analysis. J Virol. 1985 May;54(2):598–607. doi: 10.1128/jvi.54.2.598-607.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weiss R. A., Bennett P. L. Assembly of membrane glycoproteins studied by phenotypic mixing between mutants of vesicular stomatitis virus and retroviruses. Virology. 1980 Jan 30;100(2):252–274. doi: 10.1016/0042-6822(80)90518-8. [DOI] [PubMed] [Google Scholar]
- Whitt M. A., Chong L., Rose J. K. Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant. J Virol. 1989 Sep;63(9):3569–3578. doi: 10.1128/jvi.63.9.3569-3578.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wiener J. R., Pal R., Barenholz Y., Wagner R. R. Effect of the vesicular stomatitis virus matrix protein on the lateral organization of lipid bilayers containing phosphatidylglycerol: use of fluorescent phospholipid analogues. Biochemistry. 1985 Dec 17;24(26):7651–7658. doi: 10.1021/bi00347a023. [DOI] [PubMed] [Google Scholar]
- Wiener J. R., Pal R., Barenholz Y., Wagner R. R. Influence of the peripheral matrix protein of vesicular stomatitis virus on the membrane dynamics of mixed phospholipid vesicles: fluorescence studies. Biochemistry. 1983 Apr 26;22(9):2162–2170. doi: 10.1021/bi00278a017. [DOI] [PubMed] [Google Scholar]
- Wilson T., Lenard J. Interaction of wild-type and mutant M protein vesicular stomatitis virus with nucleocapsids in vitro. Biochemistry. 1981 Mar 3;20(5):1349–1354. doi: 10.1021/bi00508a048. [DOI] [PubMed] [Google Scholar]
- Zakowski J. J., Petri W. A., Jr, Wagner R. R. Role of matrix protein in assembling the membrane of vesicular stomatitis virus: reconstitution of matrix protein with negatively charged phospholipid vesicles. Biochemistry. 1981 Jun 23;20(13):3902–3907. doi: 10.1021/bi00516a037. [DOI] [PubMed] [Google Scholar]
- Zakowski J. J., Wagner R. R. Localization of membrane-associated proteins in vesicular stomatitis virus by use of hydrophobic membrane probes and cross-linking reagents. J Virol. 1980 Oct;36(1):93–102. doi: 10.1128/jvi.36.1.93-102.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van der Goot F. G., González-Mañas J. M., Lakey J. H., Pattus F. A 'molten-globule' membrane-insertion intermediate of the pore-forming domain of colicin A. Nature. 1991 Dec 5;354(6352):408–410. doi: 10.1038/354408a0. [DOI] [PubMed] [Google Scholar]