With the emergence of new viruses, such as the SARS virus and the avian influenza virus, the importance of investigations on the genetic basis of viral infections becomes clear. Sendai virus causes a localized respiratory tract infection in rodents, while a mutant, F1-R, causes a systemic infection. It has been suggested that two determinants are responsible for the systemic infection caused by F1-R [Okada et al (1998) Arch Virol 143:2343–2352]. The primary determinant of the pantropism is the enhanced proteolytic cleavability of the fusion (F) protein of F1-R, which allows the virus to undergo multiple rounds of replication in many different organs, whereas wild-type virus can only undergo multiple rounds of replication in the lungs. The enhanced cleavability of F1-R F was previously attributed to an amino acid change at F115 that is adjacent to the cleavage site at amino acid 116. Secondly, wild-type virus buds only from the apical domain of bronchial epithelium, releasing virus into the lumen of the respiratory tract, whereas F1-R buds from both apical and basolateral domains. Thus, virus is released into the basement membrane where it can easily gain access to the bloodstream for dissemination. The microtubule disruption is attributed to two amino acid differences in M protein. To confirm that the F and M gene mutations described above are solely responsible for the phenotypic differences seen in wld-type versus F1-R infections, reverse genetics was used to construct recombinant Sendai viruses with various combinations of the mutations found in the M and F genes of F1-R. Plaque assays were performed with or without trypsin addition. A recombinant virus containing all F1-R M and F mutations formed plaques in LLC-MK2 cells and underwent multiple cycles of replication without trypsin addition. To clarify which mutation(s) are necessary for plaque formation, plaque assays were done using other recombinant viruses. A virus with only the F115 change, which was previously thought to be the only change important for plaque formation of F 1-R F, did not confer upon the virus the ability to form plaques without the addition of trypsin. Another virus with the F115 and both M changes gave the same result. Therefore, more than one mutation in the F gene contributes to the ability of F1-R to form plaques without trypsin addition.
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We would like to thank Robert Desharnais, California State University, Los Angeles, for his help with the statistical analysis. Support for this research was provided by U.S. NIH MBRS SCORE grant SO6 GM 8101–30 (N.L.M.), NIH Bridges to the Ph.D. grant GM 54939 (E.S. and A.Z.), NIH MBRSRISE grant GM 61331 (A.K.), NIH Bridges to the Future grant GM 49001 (A.K.), and NIH MARC grant GM 08228 (A.K.)
Author information Authors and AffiliationsDepartment of Biological Sciences, California State University, Los Angeles, 5151 State University Drive, Los Angeles, CA, 90032, USA
Xiaogang Hou, Edgar Suquilanda, Ana Zeledon, Apollo Kacsinta, Akila Moore, Joseph Seto & Nancy McQueen
Correspondence to Nancy McQueen.
About this article Cite this articleHou, X., Suquilanda, E., Zeledon, A. et al. Mutations in Sendai virus variant F1-R that correlate with plaque formation in the absence of trypsin. Med Microbiol Immunol 194, 129–136 (2005). https://doi.org/10.1007/s00430-004-0224-3
Received: 02 February 2004
Published: 26 June 2004
Issue Date: May 2005
DOI: https://doi.org/10.1007/s00430-004-0224-3
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