ライフサイエンスコーパス: NS5

1 NS5 has 5'-RNA methyltransferase (MT)/guanylyltransferas

2 NS5 was found to be monomeric and well-folded under the

3 ver, previous studies have shown that DENV-2 NS5 accumulates in the nucleus during infection.

4 V-2 NS5, did not substantially affect DENV-2 NS5 nuclear localization, whereas knockdown of importin-

5 a isoform previously shown to bind to DENV-2 NS5, did not substantially affect DENV-2 NS5 nuclear loc

6 te the structure and dynamics of DENV type 3 NS5 in solution, we conducted small-angle X-ray scatteri

7 Interaction studies using DENV-2 and -4 NS5 and human importin-alpha isoforms failed to identify

8 ins accumulate in the nucleus, DENV-1 and -4 NS5 are predominantly if not exclusively localized to th

9 Comparative studies on the DENV-2 and -4 NS5 proteins revealed that the difference in DENV-4 NS5

10 teins revealed that the difference in DENV-4 NS5 nuclear localization was not due to rapid nuclear ex

11 he large flavivirus nonstructural protein 5 (NS5) (105 kDa) has RNA methyltransferase activities at i

12 probes, located at nonstructural protein 5 (NS5) and the 3' noncoding region (3'NC) of DENV.

13 engue virus (DENV) non-structural protein 5 (NS5) comprises an N-terminal methyltransferase and a C-t

14 Nonstructural protein 5 (NS5) contains a methyltransferase for RNA capping and a

15 DENV non-structural protein 5 (NS5) contains enzymatic activities required for capping

16 We report that nonstructural protein 5 (NS5) from four serocomplexes of flaviviruses specificall

17 sphorylation of the nonstructural protein 5 (NS5) is a conserved feature of flaviviruses, but the kin

18 Dengue virus (DENV) nonstructural protein 5 (NS5) is composed of two globular domains separated by a

19 irst time, that the nonstructural protein 5 (NS5) mediates both guanine N-7 and ribose 2'-O methylati

20 construct expressed nonstructural protein 5 (NS5), while a second recombinant expressed a soluble var

21 sitions P4-P2' surrounding the NS2-3, NS4-5, NS5-6, and NS6-7 cleavage sites contain all of the struc

22 eaved rapidly and three "late" sites (NS4-5, NS5-6, and NS6-7) processed subsequently and less effici

23 helicase and covalently linked NS3(172-618)-NS5(320-341) reveals a rigid and compact formation of th

24 Cellular immune responses against NS5 were also elicited, as evidenced by major histocompa

25 All NS5 cDNA sequences are encoded by three loci, of which t

26 All NS5 proteins inhibited HIV replication.

27 CD8 T-cell lines specific for NS3-1073 and NS5-2594 were expanded from HCV-seropositive persons by

28 observed with the addition of the NS1/2A and NS5 vaccine virus genome regions.

29 HCV proteins core, nonstructural (NS) 3, and NS5.

30 HCV antigens (core, nonstructural NS3/4 and NS5) and control phytohemagglutinin (PHA) was monitored

31 stering was similarly broad in the E1/E2 and NS5 regions.

32 unctional proteins NS3 protease/helicase and NS5 methyltransferase/RNA-dependent RNA polymerase form

33 by nonstructural protein 3 (NS3), NS2A, and NS5 were the most targeted proteins.

34 NS3 recruits NS2B and NS5 proteins to form complexes possessing protease and r

35 ntly on the capsid and nonstructural NS3 and NS5 antigens.

36 NS3 and NS5 are highly conserved among the four serotypes, and t

37 Those within NS3 and NS5 are located at the surface and/or within the NS5 dim

38 Flavivirus NS3 and NS5 are required in viral replication and 5'-capping.

39 erotype-specific interaction between NS3 and NS5 as well as specific interdomain interaction within N

40 se in IL-10 secretion in response to NS3 and NS5 in subjects with HCV compared with HIV and HCV coinf

41 the pinpointed amino acids from the NS3 and NS5 regions are also conserved.

42 ncestry with the Flaviviridae in the NS3 and NS5 regions.

43 s to nonstructural proteins 3 and 5 (NS3 and NS5).

44 core protein, nonstructural proteins NS3 and NS5, and recall antigens tetanus toxoid and Candida.

45 In DEN2-infected mammalian cells, NS3 and NS5, the viral 5'-RNA methyltransferase/polymerase, exis

46 hly conserved nonstructural proteins NS3 and NS5.

47 n each of the nonstructural proteins NS3 and NS5.

48 is required for interaction between NS3 and NS5.

49 )- alpha against HCV proteins Core, NS3, and NS5 and recall antigens.

50 ls targeting the nonstructural NS1, NS3, and NS5 proteins of TDV-2.

51 and the remaining 86, chiefly of E, NS3, and NS5, shared an identity of nine or more consecutive amin

52 s localized to antigens E1, E2, p7, NS3, and NS5.

53 E2, E2-HVR1, NS3 (helicase domain), NS4, and NS5 antigens was 97%, 98%, 28%, 88%, 33%, and 68%, respe

54 -IgG reactivities to the core, NS3, NS4, and NS5 HCV recombinant proteins and applied it to 99 serum

55 elope 2 (E2), nonstructural (NS) 3, NS4, and NS5 proteins, and NS4a and E2-HVR-1 peptides were used i

56 s from the viral core, E1, E2, NS3, NS4, and NS5 regions and different subtype-specific regions of th

57 ns: the core, E1, E2/NS1, NS2, NS3, NS4, and NS5 regions.

58 ype specific for E2, E2-HVR-1, NS3, NS4, and NS5 were detected in a minority of serum samples.

59 the HCV nonstructural proteins NS3, NS4, and NS5 will induce Ab responses, CD4+ Th cell proliferation

60 he nonstructural (NS) proteins NS3, NS4, and NS5, each of which was detected by >30% of subjects, but

61 he HCV non-structural antigens NS3, NS4, and NS5, were previously reported to induce robust and susta

62 , E2 HVR1-plus-HVR2 consensus, NS3, NS4, and NS5.

63 d among the HCV proteins core, NS3, NS4, and NS5.

64 ence of antibody reactivity to NS3, NS4, and NS5.

65 in regions encoding the NS1, NS2A, NS4A, and NS5 proteins and in the 3' untranslated region (UTR).

66 ences in the nonstructural proteins NS4b and NS5, a presumed transport protein and the viral RNA poly

67 fferences were seen in the prM, E, NS4b, and NS5 genes, while sequence differences observed within th

68 rent viral proteins (E, NS2b, NS3, NS4b, and NS5) were discovered as unique to HLA-A*0201 of infected

69 osphorylation is also shown for the NS5A and NS5 proteins, respectively, of bovine viral diarrhea vir

70 inst the HCV c-22(p), c-33(p), c-100(p), and NS5 proteins, individually or combined, but it increased

71 tantly, inhibition of JAK-STAT signaling and NS5-IFN receptor interactions were demonstrated in LGTV-

72 with binding of the viral IFN-I antagonist, NS5, to prolidase (PEPD), a cellular dipeptidase implica

73 HCV load and anti-c33c and anti-NS5 levels did not distinguish 28 HCV- and HIV-positive

74 ins immunoreacted with 62 to 93% of HCV anti-NS5-positive serum samples.

75 Because NS5 may interfere with both innate and acquired immune r

76 Examination of interactions between NS5 and cellular proteins revealed that NS5 associated w

77 and 80 nucleotides (nt), respectively, bind NS5 with similar binding affinities.

78 volutionary insights into cap-1 formation by NS5, which underlies innate immunity evasion by flavivir

79 eporter assays showed that IL-8 induction by NS5 was principally through CAAT/enhancer binding protei

80 However, the mechanisms utilized by NS5 from different flaviviruses are often quite differen

81 hree nurse shark Ig L chain isotypes, called NS5.

82 A chimeric NS5 containing the D4MT/D4GT and the D2POL domains in th

83 nctional interactions involving the chimeric NS5 protein encoded by the viral genome species is essen

84 POL activities of NS5 WT D2 and the chimeric NS5 proteins with or without the K74I mutation are simil

85 In contrast, NS5 from Kunjin virus (KUN), a naturally attenuated subt

86 ns-complementation by co-expression of WT D2 NS5 accelerated viral replication of chimeric RNA withou

87 ne mutagenesis of dengue virus type 4 (DEN4) NS5 gene generated a collection of attenuating mutations

88 The N-terminal domain of DENV NS5 has guanylyltransferase and methyltransferase (MTase

89 We show that DENV NS5 purified from Escherichia coli is a substrate for PK

90 sm of ZIKV NS5 resembles dengue virus (DENV) NS5 and not its closer relative, Spondweni virus (SPOV).

91 index and represents the first-in-class DENV-NS5 allosteric inhibitor able to target both the virus N

92 tudies showed that the substitution of DENV2 NS5 MTase or POL for DENV4 NS5 within DENV2 RNA resulted

93 he acquired mutations in the DENV2 and DENV4 NS5 MTase or POL domain or in the DENV2 NS3 helicase dom

94 titution of DENV2 NS5 MTase or POL for DENV4 NS5 within DENV2 RNA resulted in a severe attenuation of

95 C nonstructural protein 5A (NS5A) and the DV NS5 protein in CD4(+) T cells inhibit HIV replication in

96 Transfection of a plasmid expressing NS5 or a dengue virus replicon induced IL-8 gene express

97 dent RNA polymerase (RdRp) in the flaviviral NS5 protein.

98 the different strategies used by flavivirus NS5 to evade the antiviral effects of IFN-I and how this

99 Hence, flavivirus NS5 proteins exhibit a remarkable functional convergence

100 ression is decreased by all human flavivirus NS5 proteins studied.

101 ot inhibited by the expression of flavivirus NS5 protein or by YFV infection, and mumps infection did

102 two alternative conformations of flavivirus NS5 proteins.

103 as a dimer, but the functional evidence for NS5 dimer is lacking.

104 in that undergoes proteolytic processing for NS5 maturation.

105 Immune responses to HCV NS5 protein were generated by genetic immunization.

106 However, NS5 is also critical to virus replication, contributing

107 ylation in response to IFN, thus identifying NS5 as a potential IFN antagonist.

108 0 contiguous pairs of charged amino acids in NS5 were individually mutagenized to create uncharged pa

109 The serotypic differences in NS5 nuclear localization did not correlate with differen

110 time that there are serotypic differences in NS5 nuclear localization.

111 Class 2 mutations resided in NS5 (K61Q in methyltransferase and W751R in RdRp).

112 The linker region of seven residues in NS5, rich in serotype-specific residues, is important fo

113 ium concentrations and temperature influence NS5-SLA interactions in solution.

114 Mutation of a single residue in KUN NS5 to the analogous residue in WNV-NY99 NS5 (S653F) ren

115 residue in WNV-NY99 NS5 (S653F) rendered KUN NS5 an efficient inhibitor of pY-STAT1.

116 The truncated HCV RNA lacking NS5 and 3' untranslated region (3' UTR) of HCV was repli

117 report the crystal structures of full-length NS5 and its polymerase domain at 3.0 A resolution.

118 of a ternary complex between the full-length NS5 protein from dengue virus, an octameric cap-0 viral

119 there are no structural data for full-length NS5.

120 Here, we demonstrate that the LGTV NS5 JAK-STAT inhibitory domain is contained between amin

121 espite considerable separation on the linear NS5 sequence, these residues localized adjacent to each

122 ing a more distant proximity between the mAb NS5 and mAb 44.1 epitopes.

123 lation of WNV-NS5-E218A, a WNV with a mutant NS5(E218A) protein leads to survival rates and cognitive

124 The nonstructural NS5 proteins of several flaviviruses antagonize IFN-I si

125 levels of inhibition observed for mAbs NS1, NS5, CS6, and CS8.

126 core epitope regions recognized by mAbs NS2, NS5, CS6, CS8, and CS9.

127 and characterization of six mAbs (NS1, NS2, NS5, CS6, CS8, and CS9) that recognize the p22(phox) sub

128 f the DENV nonstructural proteins, with NS3, NS5, and NS1 being dominant in both donor cohorts.

129 Moreover, adenovirus-encoding core and NS3-NS5 proteins increased the secretion of bioactive TGF be

130 Here, we show by competitive NS3-NS5 interaction ELISA that the NS3 peptide spanning resi

131 In contrast, NS3-NS5 protein expression preferentially induced proinflamm

132 o genotype 1-derived HCV antigens (core, NS3-NS5) was examined in 82 patients chronically infected wi

133 nic pools that span the entire HCV core, NS3-NS5.

134 ptide spanning residues 566-585 disrupts NS3-NS5 interaction but not the null-peptide bearing the N57

135 ncoding core and nonstructural proteins (NS3-NS5) were used to express HCV proteins in HSCs.

136 R for the NS3:N570A mutant suggests that NS3-NS5 interaction plays an important role in the balanced

137 ing HCV capsid-E1-E2-NS2-NS3 and HCV NS3-NS4-NS5 in HLA-A2.1-transgenic mice.

138 vity toward either a natural substrate, NS4B-NS5 precursor, or the fluorogenic peptide substrates con

139 ra suggests that phosphorylation of the NS5A/NS5 proteins or their association with cellular kinases

140 KUN NS5 to the analogous residue in WNV-NY99 NS5 (S653F) rendered KUN NS5 an efficient inhibitor of p

141 d efficiency of N region addition (87-93% of NS5 sequences) may be a result not only of simultaneous

142 However, the MT and POL activities of NS5 WT D2 and the chimeric NS5 proteins with or without

143 acterize the stoichiometry of the complex of NS5 and SLA, and determine how solution conditions such

144 We interpret the multiple conformations of NS5 observed in solution as resulting from weak interact

145 located between the two globular domains of NS5 could be flexible.

146 Therefore, the effect of NS5 on the NS3 NTPase activity was examined.

147 also surprising given that the evolution of NS5 is restrained by the requirement to maintain functio

148 Expression of NS5 alone inhibited STAT1 phosphorylation in response to

149 Thus, the mature form of NS5, when not expressed as a precursor, was able to bind

150 mutation is associated with the function of NS5 in IFN antagonism and may influence virulence of WNV

151 udy, we examined the nuclear localization of NS5 for all four DENV serotypes.

152 ted the differential nuclear localization of NS5.

153 The varied mechanisms of NS5 as an IFN-I antagonist are also surprising given tha

154 The specific amino acid residues of NS5 involved in IFN antagonism are not known.

155 The crystal structure of NS5 shows it as a dimer, but the functional evidence for

156 Available crystal structures of NS5 fragments indicate that residues 263-271 (using the

157 1a-derived HLA-A2-restricted HCV NS3-1073 or NS5-2594 epitope were generated from a genotype 2a-deriv

158 rived from mice immunized with either NS3 or NS5 specifically lysed target cells sensitized to either

159 ecognizing the HCV nonstructural (NS) NS3 or NS5 viral peptide target were examined by mRNA transfect

160 HCV core, but not HCV E1, E2, NS3, NS4, or NS5, bound to STAT1.

161 ation of the latter, indicating that peptide NS5(320-341) engages in specific and discrete interactio

162 n containing an inactivated viral polymerase NS5 (RlucRep-NS5mt).

163 re critical for binding the viral polymerase NS5 to initiate minus-strand RNA synthesis.

164 ractions between the dengue virus polymerase NS5 and SLA in solution has not been performed.

165 The viral nonstructural protein NS5 of some flaviviruses functions as the major IFN anta

166 re, we report that the nonstructural protein NS5 of ZIKV and other flaviviruses examined could suppre

167 phalitis virus use the nonstructural protein NS5 to suppress JAK-STAT signaling.

168 rotein NS2a and two in nonstructural protein NS5, to minimize the risk of detection failure due to ge

169 of this pathway is the nonstructural protein NS5.

170 In this study, we show that the purified NS5 alone is sufficient for the synthesis of the two pro

171 To isolate the function of the viral RdRP (NS5) from that of other host or viral factors present in

172 ng on a novel dominant HLA-B*5502-restricted NS5(329-337) epitope, and assessed T-cell responses to s

173 The relatively stable NS5 region was chosen for analysis because it allowed fo

174 tional significance to the crystal structure NS5 dimer.

175 As from different serotypes, indicating that NS5 recognizes the overall shape of SLA as well as speci

176 Here, we report that NS5 from the virulent NY99 strain of WNV prevented pY-ST

177 ween NS5 and cellular proteins revealed that NS5 associated with IFN-alpha/beta and -gamma receptor c

178 The results show that NS5 nuclear localization is not strictly required for vi

179 The results show that NS5 stimulated the NS3 NTPase and RTPase activities.

180 Previous studies have shown that NS5 residue Lys-330 is required for interaction between

181 sults of these experiments also suggest that NS5 adopts multiple conformations in solution, ranging f

182 ther cellular and viral MTases suggests that NS5 requires distinct amino acids for its N-7 and 2'-O M

183 The NS5 amplimer/probe set was formulated as a one-tube, mul

184 The NS5 protocol utilizes two flaviviral consensus outer amp

185 The NS5 stimulation of NS3 NTPase was dose-dependent until a

186 The NS5 structure has striking similarities to the NS5 prote

187 sis shows that while the CTLs expressing the NS5-specific TCR reduced HCV RNA replication by a noncyt

188 ors present in the cytoplasmic extracts, the NS5 protein was expressed and purified from Escherichia

189 merase chain reaction using primers from the NS5 region of the HGV genome.

190 osorbent assay includes the protein from the NS5 region.

191 249G) together with either a mutation in the NS5 protein (A804V) or three mutations in the 3'UTR (A10

192 e was determined by direct sequencing in the NS5 region, and in the remaining 85, type-specific prime

193 acterize mechanism(s) of HIV inhibition, the NS5 proteins of GBV-C, DV, hepatitis C virus, West Nile

194 in and suggests that residues 263-268 of the NS5 protein from DENV3 are the major contributors to the

195 rus (HCV) on the antigenic properties of the NS5 protein was studied by using recombinant proteins.

196 se, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2'-

197 to compare the phosphorylation sites of the NS5 proteins of yellow fever virus (YFV) and dengue viru

198 e was determined by direct sequencing of the NS5 region of HCV with type-specific primers.

199 These unique features of the NS5-based immunoassay will be very useful for both clini

200 ze and shape of SLA and the formation of the NS5-SLA complex.

201 with ssRNA for the same binding site on the NS5 polymerase.

202 Polymerase chain reaction was done on the NS5 region and was followed by automated direct sequenci

203 ased on viral envelope and NS3 proteins, the NS5-based assay (i) reliably discriminates between WNV i

204 us antibody, and RT-PCR assays targeting the NS5 and envelope genes.

205 the C-prM was more sensitive (100%) than the NS5 (91%) or the 3'NC (91%) protocol.

206 We also observe that the NS5-SLA interaction is influenced by the magnesium conce

207 5 structure has striking similarities to the NS5 protein of the related Japanese encephalitis virus.

208 n and growth only in mice immunized with the NS5-encoding DNA construct, establishing the generation

209 are located at the surface and/or within the NS5 dimer interface, providing a functional significance

210 This NS5-STAT2 interaction requires IFN-I-induced tyrosine ph

211 The activity of this NS5 protein is verified through a de novo RdRp assay on

212 multiple nonstructural proteins ([NS] NS2 to NS5).

213 Impaired CTL responses to NS5 were corrected by syngeneic transfer of control DCs.

214 at a recombinant full-length and a truncated NS5 protein containing the methyltransferase (MTase) dom

215 lting from weak interactions between the two NS5 domains and flexibility of the linker in the absence

216 It is also unclear whether the two NS5 domains interact with each other to form a stable st

217 Dengue virus NS5 also binds SLAs from different serotypes, indicating

218 We show that dengue virus NS5 binds SLA with a 1:1 stoichiometry and that the asso

219 overed allosteric pocket on the dengue virus NS5 polymerase.

220 results indicate a role for the dengue virus NS5 protein in the induction of IL-8 by DEN2V infection.

221 Thus, the dengue virus NS5 protein inhibits HIV replication in vitro, potential

222 To determine whether the dengue virus NS5 protein inhibits HIV replication, CD4(+) T cell line

223 mall-molecule inhibitors of the dengue virus NS5 RNA capping enzyme.

224 HIV replication in dengue virus NS5-expressing cells decreased by >90% compared with con

225 Quantitatively characterizing dengue virus NS5-SLA interactions will facilitate the design and asse

226 eric inhibitor able to target both the virus NS5-NS3 interaction and the host kinases c-Src/Fyn.

227 trate here that the polymerase of the virus, NS5, binds to STAT2 and is necessary and sufficient for

228 gether to block STAT1 phosphorylation, while NS5 binds and promotes degradation of human STAT2, thus

229 e 5' termini of RNA substrates interact with NS5 during the sequential methylation reactions.

230 syngenic SP2/0 cells stably transfected with NS5.

231 l as specific interdomain interaction within NS5 required for RNA replication.

232 on prevents pY-STAT1 although a role for WNV NS5 in IFN antagonism has not been fully explored.

233 WNV-NS5-E218A-recovered mice (recovery defined as survival a

234 Hippocampi from WNV-NS5-E218A-recovered mice with poor spatial learning show

235 Inoculation of WNV-NS5-E218A, a WNV with a mutant NS5(E218A) protein leads

236 Phosphorylation of BVDV NS5A and YF NS5 was observed in infected cells, transient expression

237 ilarities in the properties of BVDV NS5A, YF NS5, and HCV NS5A phosphorylation in vitro further sugge

238 ur results demonstrate the importance of YFV NS5 in overcoming the antiviral action of IFN-I and offe

239 nique mechanism that involves binding of YFV NS5 to the IFN-activated transcription factor STAT2 only

240 at a lysine in the N-terminal region of YFV NS5.

241 hat interacts with and polyubiquitinates YFV NS5 to promote its binding to STAT2 and trigger IFN-I si

242 ZIKV NS5 expression resulted in proteasomal degradation of th

243 a high resolution structure (1.55 A) of ZIKV NS5 methyltransferase bound to a novel S-adenosylmethion

244 The mechanism of ZIKV NS5 resembles dengue virus (DENV) NS5 and not its closer

245 ion of a potential drug-binding site of ZIKV NS5, which might facilitate the development of novel ant

246 we report the crystal structure of the ZIKV NS5 protein in complex with S-adenosyl-L-homocysteine, i