Mutational analysis of the GDD sequence motif of classical swine fever virus RNA-dependent RNA polymerases.
To define the function of the GDD motif of the RNA-dependent RNA polymerase (RdRp) of classical swine fever virus (CSFV), single amino acid substitutions were introduced into the CSFV NS5B. All substitutions within the GDD motif were detrimental to the polymerase activity, the binding activity and the terminal nucleotidyl transferase activity of the NS5B protein. It was also found that the wild-type NS5B had higher RdRp activity with Mg(+2) than with Mn(+2) whereas some mutants worked better with Mn(+2) than with Mg(+2), suggesting that substitutions within the GDD motif modified the enzyme cation preferences and the GDD sequence of CSFV NS5B might be involved in polymerase-metal interaction. Therefore, the GDD amino acid sequence is important for the function of CSFV RdRp.
Wang
Yujing
Y
College of Life and Environment Sciences, Shanghai Normal University, Shanghai, 200234, China.
Xiao
Ming
M
Chen
Jun
J
Zhang
Weiqiong
W
Luo
Jianglan
J
Bao
Kan
K
Nie
Ming
M
Chen
Jiakuan
J
Li
Bo
B
D020816:Amino Acid Motifs / Q000235:genetics; D006692:Classical Swine Fever Virus / Q000235:genetics*; D004252:DNA Mutational Analysis; D005814:Genes, Viral; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D050505:Mutant Proteins / Q000378:metabolism; D011110:Polymorphism, Genetic; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics* / Q000378:metabolism; D011869:Radioligand Assay
D050505:Mutant Proteins; D008345:Manganese; D012324:RNA-Dependent RNA Polymerase; D008274:Magnesium