Restricted replication of vesicular stomatitis virus in T lymphocytes is coincident with a deficiency in a cellular protein kinase required for viral transcription
Vesicular stomatitis virus (VSV) fails to replicate in mouse T lymphocytes unless the cells have been mitogenically stimulated with concanavalin A (Con A). We have examined the possibility that the failure of VSV to replicate in unstimulated T lymphocytes can be attributed to a deficiency in a host protein kinase which activates the viral P protein by phosphorylation, thus rendering it transcriptionally competent. Soluble extracts were prepared from purified mouse T lymphocytes, with or without prior treatment with Con A. The ability of these extracts to phosphorylate bacterially synthesized P protein of two VSV serotypes was measured in vitro. Activity of the protein kinase on the P proteins of the Indiana or New Jersey serotypes of VSV increased, on average 2.4- and 2.1-fold respectively, after treatment of the cells with 3 µg/ml Con A. Higher concentrations of Con A induced proportional increases (up to 10-fold) in the activity of the host protein kinase. Activities of the kinase phosphorylating the P protein in separate populations of CD4- and CD8-containing murine T lymphocytes increased similarly on mitogenic activation. No biochemical or immunological differences were observed between the T cell protein kinase and the previously characterized protein kinase (casein kinase II) from BHK-21 cells. The activity of the kinase that phosphorylates the P protein did not vary in CV-1 cells on treatment with α- or γ-interferon, both of which inhibited VSV replication. Similarly, casein kinase II activities in Raji and SIRC cells, which do not normally support VSV growth, were the same as in BHK-21 cells. Thus restriction of VSV replication in these cells, in contrast to T lymphocytes, was not associated with a deficiency in the host casein kinase II activity.
BarikS.,
BanerjeeA. K.1991; Cloning and expression of the vesicular stomatitis virus phosphoprotein gene in Escherichia coli: analysis of phosphorylation status versus transcriptional activity. Journal of Virology 65:1719–1726
BarikS.,
BanerjeeA. K.1992a; Sequential phosphorylation of the phosphoprotein of vesicular stomatitis virus by cellular and viral protein kinases is essential for transcription activation. Journal of Virology 66:1109–1118
BarikS.,
BanerjeeA. K.1992b; Phosphorylation by cellular casein kinase II is essential for the transcriptional activity of vesicular stomatitis virus phosphoprotein P. Proceedings of the National Academy of Sciences, U.S.A. 89:6570–6574
BradfordM. M.1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Analytical Biochemistry 72:248–254
ChattopadhyayD.,
BanerjeeA. K.1987; Phosphorylation within a specific domain of the phosphoprotein of vesicular stomatitis virus regulates transcription in vitro
. Cell 49:407–414
EpsteinM. A.,
AchongB. G.,
BarrY. M.,
ZajacB.,
HenleG.,
HenleW.1966; Morphological and virological investigations on cultured Burkitt tumor lymphoblasts (strain Raji). Journal of the National Cancer Institute 37:547–551
HammondD. C.,
HaleyB. E.,
LesnawJ. A.1992; Identification and characterization of serine/threonine protein kinase activity intrinsic to the L protein of vesicular stomatitis virus New Jersey. Journal of General Virology 73:67–75
HsuC.-H.,
MorganE. M.,
KingsburyD. W.1982; Site-specific phosphorylation regulates the transcriptive activity of vesicular stomatitis virus. Journal of Biological Chemistry 260:8990–8995
JohnsonG. P.,
HermanR. C.1984; Nonpermissive infection of lymphoblastoid cells by vesicular stomatitis virus. I. Synthesis and function of the viral transcripts. Virus Research 1:259–274
JuliusM. H.,
SimpsonE.,
HerzenbergL.1973; A rapid method for the isolation of functional thymus-derived murine lymphocytes. European Journal of Immunology 3:6450649
KingsfordL.,
EmersonS. U.1980; Transcriptional activities of different phosphorylated species of NS protein purified from vesicular stomatitis virions and cytoplasm of infected cells. Journal of Virology 33:1097–1105
Muller-DeckerK.,
AmtmannE.,
SauerG.1987; Inhibition of the phosphorylation of the regulatory non-structural protein of vesicular stomatitis virus by an antiviral xanthate compound. Journal of General Virology 68:3045–3056
NowakowskiM.,
BloomB. R.,
EhrenfeldE.,
SummersD. F.1973; Restricted replication of vesicular stomatitis virus in human lymphoblastoid cells. Journal of Virology 12:1272–1278
RuddC. E.,
AndersonP.,
MorimotoC.,
StreuliM.,
SclossmanS. F.1989; Molecular interactions, T-cell subsets and a role of the CD4/CD8:p561ck complex in human T cell activation. Immunological Reviews 111:225–266
SanchezA.,
DeB. P.,
BanerjeeA. K.1985; In vitro phosphorylation of NS protein by the L protein of vesicular stomatitis virus. Journal of General Virology 66:1025–1036
SauerG.,
AmtmannE.,
MelberK.,
KnappA.,
MullerK.,
HummelK.,
SchermA.1984; DNA and RNA virus species are inhibited by xanthates, a class of antiviral compounds with unique properties. Proceedings of the National Academy of Sciences, U.S.A. 81:3263–3267
SinacoreM. S.,
Lucas-LenardJ.1982; The effect of the vesicular stomatitis virus-associated protein kinase on mRNA transcription in vitro
. Virology 121:404–413
ThacoreH. R.,
YoungnerJ. S.1975; Abortive infection of a rabbit cornea cell line by vesicular stomatitis virus: conversion of productive infection by superinfection with vaccinia virus. Journal of Virology 16:322–329
TowbinH.,
StaehelinT.,
GordonJ.1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proceedings of the National Academy of Sciences, U.S.A. 76:4350–4354
WebbD. R.,
MunshiS.,
BanerjeeA. K.1981; Replication of vesicular stomatitis virus in murine spleen cells: enrichment of the virus-replicating lymphocytes and analysis of replication restriction. Infection and Immunity 32:169–172
WethersJ. A.,
JohnsonG. P.,
SchumacherC. L.,
HermanR. C.1985; Nonpermissive infection of lymphoblastoid cells by vesicular stomatitis virus. II. Effect on viral morphogenesis. Virus Research 2:345–358
WinterE.,
YamamotoF.,
AlmogueraC.,
PeruchoM.1985; A method to detect and characterize point mutations in transcribed genes: amplification and over-production of the mutant c-Ki-ras allele in human tumor cells. Proceedings of the National Academy of Sciences, U.S.A. 75:4257–4261
Restricted replication of vesicular stomatitis virus in T lymphocytes is coincident with a deficiency in a cellular protein kinase required for viral transcription