Sindbis virus mutations which coordinately affect glycoprotein processing, penetration, and virulence in mice.
Rapid penetration of baby hamster kidney cells was used as a selective pressure for the isolation of pathogenesis mutants of the S.A.AR86 strain of Sindbis virus. Unlike most Sindbis virus strains, S.A.AR86 is virulent in adult as well as neonatal mice. Two classes of mutants were defined. One class was attenuated in adult mice inoculated intracerebrally as well as in neonatal mice inoculated either intracerebrally or subcutaneously. Sequence analysis of the glycoprotein genes of the parent virus and three such mutant strains revealed a single point mutation which resulted in an amino acid change at position 1 in the E2 glycoprotein. The change from a serine in S.A.AR86 to an asparagine in the mutants created a new site for N-linked glycosylation which appeared to be utilized. This mutation did not retard release of infectious particles; however, mutant virions contained the E2 precursor protein (PE2) rather than the E2 glycoprotein itself. The mutants also lost the ability to bind two E2-specific monoclonal antibodies, R6 and R13. A second class of mutants was attenuated in neonatal mice upon subcutaneous inoculation but remained virulent in adults and in neonates when inoculated intracerebrally. Sequence analysis of three such strains revealed the substitution of an arginine residue for a serine at position 114 in the E2 glycoprotein. Reactivity with monoclonal antibodies R6 and R13 was reduced, yet members of this mutant class were more susceptible than S.A.AR86 to neutralization by these antibodies.
Russell
D L
DL
Department of Microbiology, North Carolina State University, Raleigh 27695-7615.
Dalrymple
J M
JM
Johnston
R E
RE
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn / Q000382:microbiology; D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009422:Nervous System Diseases / Q000382:microbiology*; D012367:RNA, Viral / Q000235:genetics; D012845:Sindbis Virus / Q000235:genetics* / Q000472:pathogenicity; D013329:Structure-Activity Relationship; D014759:Viral Envelope Proteins / Q000235:genetics*; D014779:Virus Replication
D000956:Antigens, Viral; D008562:Membrane Glycoproteins; D012367:RNA, Viral; D014759:Viral Envelope Proteins