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Journal of General Virology (2000), 81, 161-170.
© 2000 Society for General Microbiology


Animal: RNA Viruses

Linkage of an alphavirus host-range restriction to the carbohydrate-processing phenotypes of the host cell

Karl W. Boehme1, Jacqueline C. Williams1, Robert E. Johnston2 and Hans W. Heidner1

Division of Life Sciences, University of Texas at San Antonio, 6900 North Loop 1604 West, San Antonio, TX 78249-0662, USA1
Department of Microbiology and Immunology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599-7290, USA2

Author for correspondence: Hans Heidner. Fax +1 210 458 5658. e-mail hheidner{at}utsa.edu

The Sindbis virus mutant NE2G216 retains PE2 in place of E2 in its virion structure. NE2G216 is a host-range mutant that replicates with near-normal kinetics in vertebrate cells, but displays severely restricted growth in cultured mosquito cells (C6/36) due to defects in the virus maturation process. In this study we tested the hypothesis that the host-range phenotype of NE2G216 was linked to the differences in carbohydrate-processing phenotypes between vertebrate and arthropod cells. Arthropod cell-derived glycoproteins are distinguishable from those synthesized in vertebrate cells by the absence of complex- and hybrid-type N-linked oligosaccharides. To test our hypothesis we compared the growth of the wild-type virus, TRSB, NE2G216 and three PE2-containing, C6/36 cell-adapted variants, in vertebrate cells treated with 1-deoxymannojirimycin (1-dMM). 1-dMM inhibits the Golgi {alpha}-mannosidase I enzyme and limits oligosaccharide processing to high-mannose forms (Man8–9GlcNAc2). The growth of TRSB was not restricted by the action of 1-dMM; however, NE2G216 was restricted in a dose-dependent manner. In contrast, the growth of each PE2-containing, C6/36 cell-adapted mutant was enhanced by low concentrations of 1-dMM (up to 1500%) and was only slightly affected by the higher concentrations. These results demonstrate that virion maturation functions of NE2G216 are sensitive to the structure of cis-linked oligosaccharides, and indicate that the carbohydrate-processing phenotypes of the host cell can influence viral host-range and function as a selective pressure in alphavirus evolution.




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