TY - JOUR
T1 - Two mutations in the C-terminal domain of influenza virus RNA polymerase PB2 enhance transcription by enhancing cap-1 RNA binding activity
AU - Zhang, Shijian
AU - Wang, Qiang
AU - Wang, Jinlan
AU - Mizumoto, Kiyohisa
AU - Toyoda, Tetsuya
N1 - Funding Information:
The sequences of the primers used in this study are available from the authors upon request. This work was supported by Grants-in-Aids from the Chinese Academy of Sciences ( 0514P51131 ), National Science Foundation of China ( 30670090 and 30970153 ), Li Ka Shing Foundation ( 0682P11131 ), RESPARI ( 0581P14131 ), and FLUINNATE ( SP5B-CT-2006-044161 ).
Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.
PY - 2012/1
Y1 - 2012/1
N2 - Influenza virus RNA polymerase (RdRp) PB2 is the cap-1 binding subunit and determines host range and pathogenicity. The mutant human influenza virus RdRp containing PB2 D701N and D701N/S714R demonstrated enhanced replicon activity in mammalian cells. We investigated the influence of these mutations on RdRp activity. Cap-1-dependent transcription activities of D701N/S714R, D701N, and S714R were 348.1 ± 6.2%, 146.4 ± 11%, and 250.1 ± 0.8% of that of the wild type (wt), respectively. Replication activity of these mutants for complimentary RNA to viral RNA ranged from 44% to 53% of that of the wt. Cap-1 RNA-binding activities of D701N/S714R, D701N, and S714R were 262 ± 25%, 257 ± 34%, and 315 ± 9.6% of that of the wt, respectively, and their cap-dependent endonuclease activities were similar to that of the wt. These mutations did not affect template RNA-binding activities. D701N and S714R mutations enhanced transcription by enhancing cap-1 RNA-binding activity, but they may exhibit decreased efficiency of priming by the cap-1 primer. These mutations at the C-terminal domain of PB2 may affect its cap-binding domain.
AB - Influenza virus RNA polymerase (RdRp) PB2 is the cap-1 binding subunit and determines host range and pathogenicity. The mutant human influenza virus RdRp containing PB2 D701N and D701N/S714R demonstrated enhanced replicon activity in mammalian cells. We investigated the influence of these mutations on RdRp activity. Cap-1-dependent transcription activities of D701N/S714R, D701N, and S714R were 348.1 ± 6.2%, 146.4 ± 11%, and 250.1 ± 0.8% of that of the wild type (wt), respectively. Replication activity of these mutants for complimentary RNA to viral RNA ranged from 44% to 53% of that of the wt. Cap-1 RNA-binding activities of D701N/S714R, D701N, and S714R were 262 ± 25%, 257 ± 34%, and 315 ± 9.6% of that of the wt, respectively, and their cap-dependent endonuclease activities were similar to that of the wt. These mutations did not affect template RNA-binding activities. D701N and S714R mutations enhanced transcription by enhancing cap-1 RNA-binding activity, but they may exhibit decreased efficiency of priming by the cap-1 primer. These mutations at the C-terminal domain of PB2 may affect its cap-binding domain.
KW - Cap-1 RNA binding
KW - Cap-dependent nuclease
KW - Influenza virus
KW - PB2
KW - RNA polymerase
KW - Transcription
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U2 - 10.1016/j.bbagrm.2011.11.006
DO - 10.1016/j.bbagrm.2011.11.006
M3 - Article
C2 - 22146492
AN - SCOPUS:84055224044
VL - 1819
SP - 78
EP - 83
JO - Biochimica et Biophysica Acta - Gene Regulatory Mechanisms
JF - Biochimica et Biophysica Acta - Gene Regulatory Mechanisms
SN - 1874-9399
IS - 1
ER -