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Journal of Virology, May 2000, p. 4220-4228, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Single Deletion in the Membrane-Proximal Region of the Sindbis Virus Glycoprotein E2 Endodomain Blocks Virus Assembly

Raquel Hernandez,1 Heuiran Lee,2 Christine Nelson,1 and Dennis T. Brown1,*

Department of Biochemistry, North Carolina State University, Raleigh, North Carolina 27695,1 and Yonsei Cancer Center, Institute of Cancer Research, Yonsei University College of Medicine, Seoul, Korea2

Received 18 November 1999/Accepted 7 February 2000

The envelopment of the Sindbis virus nucleocapsid in the modified cell plasma membrane involves a highly specific interaction between the capsid (C) protein and the endodomain of the E2 glycoprotein. We have previously identified a domain of the Sindbis virus C protein involved in binding to the E2 endodomain (H. Lee and D. T. Brown, Virology 202:390-400, 1994). The C-E2 binding domain resides in a hydrophobic cleft with C Y180 and W247 on opposing sides of the cleft. Structural modeling studies indicate that the E2 domain, which is proposed to bind the C protein (E2 398T, 399P, and 400Y), is located at a sufficient distance from the membrane to occupy the C protein binding cleft (S. Lee, K. E. Owen, H. K. Choi, H. Lee, G. Lu, G. Wengler, D. T. Brown, M. G. Rossmann, and R. J. Kuhn, Structure 4:531-541, 1996). To measure the critical spanning length of the E2 endodomain which positions the TPY domain into the putative C binding cleft, we have constructed a deletion mutant, Delta K391, in which a nonconserved lysine (E2 K391) at the membrane-cytoplasm junction of the E2 tail has been deleted. This mutant was found to produce very low levels of virus from BHK-21 cells due to a defect in an unidentified step in nucleocapsid binding to the E2 endodomain. In contrast, Delta K391 produced wild-type levels of virus from tissue-cultured mosquito cells. We propose that the phenotypic differences displayed by this mutant in the two diverse host cells arise from fundamental differences in the lipid composition of the insect cell membranes which affect the physical and structural properties of membranes and thereby virus assembly. The data suggest that these viruses have evolved properties adapted specifically for assembly in the diverse hosts in which they grow.

* Corresponding author. Mailing address: Department of Biochemistry, North Carolina State University, Campus Box 7622, Raleigh, NC 27695-7622. Phone: (919) 515-5802. Fax: (919) 515-2047. E-mail: dennis_brown{at}ncsu.edu.

Journal of Virology, May 2000, p. 4220-4228, Vol. 74, No. 9
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Hafer, A., Whittlesey, R., Brown, D. T., Hernandez, R. (2009). Differential Incorporation of Cholesterol by Sindbis Virus Grown in Mammalian or Insect Cells. J. Virol. 83: 9113-9121 [Abstract] [Full Text]  
  • Whitehurst, C. B., Soderblom, E. J., West, M. L., Hernandez, R., Goshe, M. B., Brown, D. T. (2007). Location and Role of Free Cysteinyl Residues in the Sindbis Virus E1 and E2 Glycoproteins. J. Virol. 81: 6231-6240 [Abstract] [Full Text]  
  • West, J., Hernandez, R., Ferreira, D., Brown, D. T. (2006). Mutations in the Endodomain of Sindbis Virus Glycoprotein E2 Define Sequences Critical for Virus Assembly. J. Virol. 80: 4458-4468 [Abstract] [Full Text]  
  • West, J., Brown, D. T. (2006). Role of a conserved tripeptide in the endodomain of Sindbis virus glycoprotein E2 in virus assembly and function.. J. Gen. Virol. 87: 657-664 [Abstract] [Full Text]  
  • Hernandez, R., Ferreira, D., Sinodis, C., Litton, K., Brown, D. T. (2005). Single Amino Acid Insertions at the Junction of the Sindbis Virus E2 Transmembrane Domain and Endodomain Disrupt Virus Envelopment and Alter Infectivity. J. Virol. 79: 7682-7697 [Abstract] [Full Text]  
  • Hernandez, R., Sinodis, C., Horton, M., Ferreira, D., Yang, C., Brown, D. T. (2003). Deletions in the Transmembrane Domain of a Sindbis Virus Glycoprotein Alter Virus Infectivity, Stability, and Host Range. J. Virol. 77: 12710-12719 [Abstract] [Full Text]  


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