Cauliflower mosaic virus ORF III product forms a tetramer in planta: its implication in viral DNA folding during encapsidation.
Cauliflower mosaic virus (CaMV) open reading frame (ORF) III encodes a 15 kDa protein; the function of which is as yet unknown. This protein has non-sequence-specific DNA binding activity and is associated with viral particles, suggesting that the ORF III product (P3) is involved in the folding of CaMV DNA during encapsidation. In this study, we demonstrated that P3 forms a tetramer in CaMV-infected plants. A P3-related protein with an apparent molecular weight of 60 kDa was detected by Western blotting analysis using anti-P3 antiserum under non-reducing conditions, while only 15 kDa P3 was detected under reducing conditions. Analysis of P3 using viable mutants with a 27-bp insertion in either ORF III or IV revealed that the 60 kDa protein was a tetramer of P3. The P3 tetramer co-sedimented with viral coat protein in multiple fractions on sucrose gradient centrifugation, suggesting that P3 tetramer binds to mature and immature virions. These results strongly suggested that CaMV P3 forms a tetramer in planta and that disulfide bonds are involved in its formation and/or stabilization. The finding of P3 tetramer in planta suggested that viral DNA would be folded compactly by the interaction with multiple P3 molecules, which would form tetramers, while being packaged into the capsid shell.
Tsuge
S
S
Laboratory of Plant Pathology, Faculty of Agriculture, Kyoto Prefectural University, Japan. s_tsuge@kpu.ac.jp
Kobayashi
K
K
Nakayashiki
H
H
Mise
K
K
Furusawa
I
I
D001937:Brassica / Q000821:virology; D002213:Capsid / Q000378:metabolism; D017796:Caulimovirus / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D002458:Cell Fractionation; D004261:DNA Replication; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames; D014764:Viral Proteins / Q000235:genetics / Q000502:physiology*; D019065:Virus Assembly; D014779:Virus Replication
D004279:DNA, Viral; D014764:Viral Proteins; C115486:p15 protein, Cauliflower mosaic virus