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      Nuclear-export-signal-dependent protein translocation of dUTPase encoded by Singapore grouper iridovirus.

      Archives of Virology
      Amino Acid Sequence, Animals, Cell Line, Cloning, Molecular, Computational Biology, Fishes, Gene Expression Profiling, Gene Expression Regulation, Viral, physiology, Iridovirus, genetics, metabolism, Molecular Sequence Data, Pyrophosphatases, Sequence Alignment, Viral Proteins, chemistry

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          Abstract

          The dUTPase is a ubiquitous and crucial enzyme responsible for regulating cellular levels of dUTP. In the present study, the expression pattern and translocation of a dUTPase homolog encoded by Singapore grouper iridovirus (SGIV) were elucidated. The SGIV ORF049R encodes a dUTPase homolog, which is a peptide of 155 amino acids that contains five conserved motifs. The temporal expression pattern during infection in vitro revealed that the SGIV dUTPase was an early transcript. A leucine-rich nuclear export signal (NES) at the C-terminus was predicted using CBS Online Servers. Subcellular location analysis showed that SGIV dUTPase is a cytoplasmic protein. Site-direct mutagenesis by overlap extension-PCR indicated that the NES is crucial for the translocation of SGIV dUTPase from the nucleus to the cytoplasm. We have discovered for the first time that the NES-dependent translocation of dUTPase is different for SGIV than for members of other species, which depend on a nuclear localization signal. These results provide new insights into the pathogenesis of fish iridoviruses.

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