The 42-kDa coat protein of Andean potato mottle virus acts as a transcriptional activator in yeast.
Interactions of viral proteins play an important role in the virus life cycle, especially in capsid assembly. Andean potato mottle comovirus (APMoV) is a plant RNA virus with a virion formed by two coat proteins (CP42 and CP22). Both APMoV coat protein open reading frames were cloned into pGBT9 and pGAD10, two-hybrid system vectors. HF7c yeast cells transformed with the p9CP42 construct grew on yeast dropout selection media lacking tryptophan and histidine. Clones also exhibited beta-galactosidase activity in both qualitative and quantitative assays. These results suggest that CP42 protein contains an amino acid motif able to activate transcription of His3 and lacZ reporter genes in Saccharomyces cerevisiae. Several deletions of the CP42 gene were cloned into the pGBT9 vector to locate the region involved in this activation. CP42 constructions lacking 12 residues from the C-terminal region and another one with 267 residues deleted from the N-terminus are still able to activate transcription of reporter genes. However, transcription activation was not observed with construction p9CP42deltaC57, which does not contain the last 57 amino acid residues. These results demonstrate that a transcription activation domain is present at the C-terminus of CP42 between residues 267 and 374.
Vidal
M S
MS
Laboratório de Genética Molecular Vegetal, Departamento de Genética, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
Margis
R
R
D001483:Base Sequence; D002213:Capsid / Q000502:physiology*; D017798:Comovirus / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000821:virology*; D011198:Solanum tuberosum / Q000821:virology*; D015533:Transcriptional Activation / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D019065:Virus Assembly
D017931:DNA Primers; D018744:DNA, Plant