Isolation and enzymatic characterization of protein lambda 2, the reovirus guanylyltransferase.
Protein lambda 2 of reovirus serotype 3 has been purified to homogeneity from extracts of cells infected with hybrid vaccinia virus strain WR into whose TK gene of the reovirus L2 genome segment under the control of the CPV ATI protein gene promoter had been inserted. Protein lambda 2 is formed in large amounts (final purification factor about 180) as a monomer that shows no tendency to pentamerize into the reovirus core projections/spikes. Isolated protein lambda 2 is reversibly guanylylated by GTP (that is, it carries out the GTP-PPi exchange reaction) and can transfer the -GMP moiety to GTP to form GppppG, to GDP to form GpppG, and to 5'-pp-terminated RNA to form GpppG- caps. These studies confirm previous studies on reovirus cores that indicated that protein lambda 2 is the reovirus guanylyltransferase. Protein lambda 2 possesses neither nucleoside nor RNA triphosphatase activities, nor methyltransferase activities; thus it is the reovirus capping enzyme, but provides neither the required 5'-ppG-terminated substrate nor does it methylate the cap structure. These must be functions of lambda 2 pentamers or of other individual or complexed components of reovirus cores.
Mao
Z X
ZX
Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina 27710.
Joklik
W K
WK
D000818:Animals; D002852:Chromatography, Ion Exchange; D011756:Diphosphates / Q000378:metabolism; D005822:Genetic Vectors; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D007739:L Cells; D012089:Mammalian orthoreovirus 3 / Q000201:enzymology* / Q000235:genetics; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D009713:Nucleotidyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics; D014758:Viral Core Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D011756:Diphosphates; D012333:RNA, Messenger; D014758:Viral Core Proteins; C071342:reovirus protein lambda2; D006160:Guanosine Triphosphate; D008715:Methionine; D013937:Thymidine Kinase; D009713:Nucleotidyltransferases; C020842:guanylyltransferase