Phosphorylation of the termini of Cauliflower mosaic virus precapsid protein is important for productive infection.
Cauliflower mosaic virus (CaMV) coat protein precursor (pre-CP) has 489 amino acids (p57) and is processed by the viral proteinase into three major forms: p44, p39, and p37. The N- and C-terminal extensions of pre-CP are released during maturation by the virus-encoded proteinase. We showed that these extensions are phosphorylated at several sites by host casein kinase II (CKII). We have identified the phosphorylated amino acids using an in vitro phosphorylation assay and tested the effect of mutation of these sites on viral infectivity. Mutation of serines S66, S68, and S72 to alanine in the N-terminal extension abolished phosphorylation of the protein in vitro. Also, mutation of all S and T residues in the C-terminus (450 to 489) made this region insensitive to CKII. Amino acid substitutions also were introduced into a full-length infectious clone of CaMV. Mutated forms of the virus with S66, S68, and S72 substituted with A or D showed a delay in symptom development and affected the infectivity of the virus. However, a mutant with an A substitution of all the S and T residues of the C-terminal extension of CP was not infectious. These results suggest that phosphorylation of the N- and C-termini of CaMV pre-CP plays an important role in the initiation of viral infection.
Champagne
Julie
J
Centre de Recherche en Infectiologie, CHUQ, Pavilion CHUL, Québec G1V 4G2, Canada.
Laliberté-Gagné
Marie-Eve
ME
Leclerc
Denis
D
D000595:Amino Acid Sequence; D001937:Brassica / Q000821:virology; D036022:Capsid Proteins / Q000737:chemistry* / Q000378:metabolism*; D047390:Casein Kinase II / Q000378:metabolism; D017796:Caulimovirus / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D002999:Clone Cells; D004279:DNA, Viral / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010766:Phosphorylation; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism
D036022:Capsid Proteins; D004279:DNA, Viral; D050505:Mutant Proteins; D011498:Protein Precursors; D011994:Recombinant Proteins; D047390:Casein Kinase II