Structural Basis of the Influenza A Virus RNA Polymerase PB2 RNA-binding Domain Containing the Pathogenicity-determinant Lysine 627 Residue

被引:79
作者
Kuzuhara, Takashi [1 ]
Kise, Daisuke [1 ]
Yoshida, Hiroko [1 ]
Horita, Takahiro [1 ]
Murazaki, Yoshimi [1 ]
Nishimura, Akie [1 ]
Echigo, Noriko [1 ]
Utsunomiya, Hiroko [2 ]
Tsuge, Hideaki [2 ]
机构
[1] Tokushima Bunri Univ, Fac Pharmaceut Sci, Biochem Lab, Tokushima 7708514, Japan
[2] Tokushima Bunri Univ, Inst Hlth Sci, Tokushima 7708514, Japan
关键词
AMINO-ACID; SUBUNIT; NUCLEOPROTEIN; OSELTAMIVIR; PROMOTER; PROGRAM; PROTEIN;
D O I
10.1074/jbc.C800224200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Because the influenza A virus has an RNA genome, its RNA-dependent RNA polymerase, comprising the PA, PB1, and PB2 subunits, is essential for viral transcription and replication. The binding of RNAprimers/promoters to the polymerases is an initiation step in viral transcription. In our current study, we reveal the 2.7 A tertiary structure of the C-terminal RNA-binding domain of PB2 by x-ray crystallography. This domain incorporates lysine 627 of PB2, and this residue is associated with the high pathogenicity and host range restriction of influenza A virus. We found from our current analyses that this lysine is located in a unique "phi"-shaped structure consisting of a helix and an encircled loop within the PB2 domain. By electrostatic analysis, we identified a highly basic groove along with this phi loop and found that lysine 627 is located in the phi loop. A PB2 domain mutant in which glutamic acid is substituted at position 627 shows significantly lower RNA binding activity. This is the first report to show a relationship between RNA binding activity and the pathogenicity-determinant lysine 627. Using the Matras program for protein three-dimensional structural comparisons, we further found that the helix bundles in the PB2 domain are similar to that of activator 1, the 40-kDa subunit of DNA replication clamp loader (replication factor C), which is also an RNA-binding protein. This suggests a functional and structural relationship between the RNA-binding mechanisms underlying both influenza A viral transcription and cellular DNA replication. Our present results thus provide important new information for developing novel drugs that target the primer/promoter RNA binding of viral RNA polymerases.
引用
收藏
页码:6855 / 6860
页数:6
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