1993 Fiscal Year Final Research Report Summary
ANALYSIS OF FUNCTIONAL PROPERTIES OF VIRAL
| Project/Area Number |
04670269
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Virology
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| Research Institution | University of Tokyo |
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Principal Investigator |
IWAMOTO Aikichi UNIVERSITY OF TOKYO, MEDICINE, ASSOCIATE PROFESSOR, 医学部(医), 助教授 (10133076)
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| Co-Investigator(Kenkyū-buntansha) |
YANAGI Yusuke UNIVERSITY OF TOKYO, MEDICINE, ASSISTANT PROFESSOR, 医学部(医), 助手 (40182365)
YOSHIKURA Hiroshi UNIVERSITY OF TOKYO, MEDICINE, PROFESSOR, 医学部(医), 教授 (60012754)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Measles virus / Retrovirus / Viral structural protein / Gag protein / Env protein |
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| Research Abstract |
In order to examine the functional properties of a component of the viral structural proteins, we introduced the gene encoding the component into the cells and analyzed them.
We subcloned Measles virus glycoprotein H and F, respectively, into a BPV vector carrying neomycin resistance gene. We introduced these plasmids into human T cell line, Jarkat. H and F were hardly exprressed on the cell surface. We then used stronger expression vector, pCXN2 with CMV enhancer and chicken beta-globin promoter and neomycin resistance gene. We introduced the plasmids into Vero cells and looked for the clones showing cell surface expression using fluorescent antibodies. We could find clones expressing H but not those expressing F.The Vero cell clone expressing H and its parental Vero cells were not different in susceptibility to measles virus infection. Therefore, it appears that the cell surface expression of H protein which can bind to measles virus receptor is not sufficient to cause interference with viral infection.
Our initial attempt to express murine leukemia virus (MLV) capsid protein (CA) in NIH3T3 cells led us to postulate a possible nuclear localization signal. However, subsequent experiments using stronger promters have been unconvincing. Even the strongest promoter could not provide a high expression of CA.A cryptic sprising-donor sequence in CA might hinder the RNA expression. We found that a mutant Env protein of Friend MLV with a substitution of 361st amino acid from Cys to Arg accumulates in ER and can function as a dominant negative mutant upon a MLV superinfection without expression on the cell surface.
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[Publications] Hijikata, M., Shimizu, Y.K., Kato, H., Iwamoto, A., Shih, J.W., Alter, H.J., Purcel, R.H., and Yoshikura, H.: "Equilibrium centrifugation studies of hepatitis C virus : evidence for circulating immune complexes." J.Virology. vol.67. 1953-1958 (1993)
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