Ribonucleotide reductase of shrimp white spot syndrome virus (WSSV): expression and enzymatic activity in a baculovirus/insect cell system and WSSV-infected shrimp.
Infection of shrimp cells with white spot syndrome virus (WSSV) results in an increase in ribonucleotide reductase (RR) expression at the RNA level. In this article we further express and characterize the induction of a novel ribonucleotide reductase after WSSV infection of shrimp cells. A baculovirus/insect system was used to express the two recombinant protein subunits RR1 and RR2, and a DNA polymerase coupled RR activity assay showed a marked increase in ribonucleotide reductase activity when cell extracts containing recombinant RR1 and RR2 were combined. The same assay revealed that RR activity increased as infection advanced in the gills of experimentally infected shrimp. An increase in RR expression was also detected at the protein level in WSSV-infected shrimp cells. An immunocytochemistry assay by confocal laser scanning microscopy showed that in hemocytes collected from WSSV-infected shrimp, both of the subunit proteins (RR1 and RR2) were concentrated mainly around the nucleus, but only RR1 was detected inside it. All of these results suggest that WSSV RR is functionally involved during WSSV infection.
Lin
Shinn-Tsuen
ST
Department of Zoology, National Taiwan University, Taipei, Taiwan, Republic of China.
Chang
Yun-Shiang
YS
Wang
Han-Ching
HC
Tzeng
Huey-Fen
HF
Chang
Zee-Fen
ZF
Lin
Jung-Yaw
JY
Wang
Chung-Hsiung
CH
Lo
Chu-Fang
CF
Kou
Guang-Hsiung
GH
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D004267:DNA Viruses / Q000201:enzymology*; D033364:Decapoda / Q000821:virology*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000096:biosynthesis; D012264:Ribonucleotide Reductases / Q000032:analysis / Q000378:metabolism*; D018411:Spodoptera; D013270:Stomach / Q000201:enzymology / Q000821:virology
D011994:Recombinant Proteins; D012264:Ribonucleotide Reductases