Influenza virus RNA polymerase PA subunit is a novel serine protease with Ser624 at the active site

被引:60
作者
Hara, K
Shiota, M
Kido, H
Ohtsu, Y
Kashiwagi, T
Iwahashi, J
Hamada, N
Mizoue, K
Tsumura, N
Kato, H
Toyoda, T [1 ]
机构
[1] Kurume Univ, Sch Med, Dept Virol, Kurume, Fukuoka 8300011, Japan
[2] Kurume Univ, Sch Med, Dept Pediat, Kurume, Fukuoka 8300011, Japan
[3] Univ Tokushima, Inst Enzyme Res, Div Enzyme Chem, Tokushima 7708503, Japan
[4] Taisho Pharmaceut Co Ltd, Med Res Labs, Omiya, Saitama 3308530, Japan
关键词
D O I
10.1046/j.1365-2443.2001.00399.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Background: Influenza virus RNA polymerase is a multifunctional enzyme that catalyses both transcription and replication of the RNA genome. The function of the influenza virus RNA polymerase PA subunit in viral replication is poorly understood, although the enzyme is known to be required for cRNA --> vRNA synthesis. The protease related activity of PA has been discussed ever since protease-inducing activity was demonstrated in transfection experiments. Results: PA protein was highly purified from insect cells infected with the recombinant baculovirus carrying PA cDNA, and a novel chymotrypsin-type serine protease activity was identified with the synthetic peptide, Suc-LLVY-MCA, in the PA protein. [H-3]DFP was crosslinked with PA and a mutational analysis revealed that serine624 was as an active site for the protease activity. Conclusions:These results constitute the demonstration of protease activity in PA subunit of the influenza virus RNA polymerase complexes.
引用
收藏
页码:87 / 97
页数:11
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