Identification and characterization of serine/threonine protein kinase activity intrinsic to the L protein of vesicular stomatitis virus New Jersey
1 School of Biological Sciences and 2 Department of Biochemistry, Lucille P. Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0225, U.S.A. A photoaffinity analogue of ATP, 8-azido-adenosine 5'-triphosphate (8-N 3 ATP), was used to probe ATP-binding sites in native transcr...
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Veröffentlicht in: | Journal of general virology 1992-01, Vol.73 (1), p.67-75 |
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Sprache: | eng |
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Zusammenfassung: | 1 School of Biological Sciences
and 2 Department of Biochemistry, Lucille P. Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0225, U.S.A.
A photoaffinity analogue of ATP, 8-azido-adenosine 5'-triphosphate (8-N 3 ATP), was used to probe ATP-binding sites in native transcription complexes of vesicular stomatitis virus (VSV) (New Jersey serotype). The analogue was found to be a substrate for a serine/threonine protein kinase that phosphorylated both the NS and L proteins of native complexes. The analogue failed to interact with the RNA polymerase, another ATP-utilizing activity associated with the transcription complex. Kinetic analyses of both ATP and 8-N 3 ATP utilization by the protein kinase yielded biphasic saturation curves. Photolysis of 8-N 3 ATP in the presence of VSV transcription complexes resulted in selective labelling of the L protein. The photolabelling of L was saturable and apparently biphasic. Photolabelling of the L protein was significantly reduced by competition with ATP whereas other nucleoside triphosphates (GTP, UTP and CTP) were ineffective competitors. The stoichiometry of photolabelling was 0.2 at 10 µ M -8N 3 ATP and 1.3 at 100 µ M -ATP. These data provide chemical evidence for a virus-encoded serine/threonine protein kinase which resides on the L protein.
Received 10 May 1991;
accepted 30 September 1991. |
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ISSN: | 0022-1317 1465-2099 |
DOI: | 10.1099/0022-1317-73-1-67 |