ライフサイエンスコーパス: tRNA(Gln

1 approximately one order of magnitude towards tRNA(Gln).

2 enzyme is rate-limiting for synthesis of Gln-tRNA(Gln).

3 ential for the production of misacylated Glu-tRNA(Gln).

4 ynthesizes Glu-tRNA(Glu) and cannot make Glu-tRNA(Gln).

5 robustly aminoacylates tRNA(Glu1) instead of tRNA(Gln).

6 of ATP hydrolysis requires both Gln and Glu-tRNA(Gln).

7 e level as ATP, but without formation of Gln-tRNA(Gln).

8 ylation of mt-tRNA(Glu), mt-tRNA(Lys) and mt-tRNA(Gln).

9 n via the invariant 3'-terminal adenosine of tRNA(Gln).

10 the synthesis of both Asn-tRNA(Asn) and Gln-tRNA(Gln).

11 idotransferase converts Glu-tRNA(Gln) to Gln-tRNA(Gln).

12 GluRS2 specifically aminoacylating Glu onto tRNA(Gln).

13 the GatCAB amidotransferase, which forms Gln-tRNA(Gln).

14 g the misaminoacylated tRNA intermediate Glu-tRNA(Gln).

15 apable of forming both Glu-tRNA(Glu) and Glu-tRNA(Gln).

16 e GatDE converts this mischarged tRNA to Gln-tRNA(Gln).

17 ) in mitochondrial tRNA(Lys), tRNA(Glu), and tRNA(Gln).

18 luRS2 only misacylates tRNA(Gln) to form Glu-tRNA(Gln).

19 otransferase to convert Glu-tRNA(Gln) to Gln-tRNA(Gln).

20 le to transamidate M. thermautotrophicus Glu-tRNA(Gln).

21 s and use either tRNAGlu or both tRNAGlu and tRNAGln.

22 AGlu, whereas GluRS2 was specific solely for tRNAGln.

23 and in E. coli, but does not charge E. coli tRNAGln.

24 ions containing [4-thio]-uridine-labeled pre-tRNAGln.

25 AGln but by a specific transamidation of Glu-tRNAGln.

26 ficity despite making no hydrogen bonds with tRNAGln.

27 ously been determined for the nucleotides in tRNAGln.

28 sequence-specific contacts between GlnRS and tRNAGln.

29 ognate tRNAs but to a decreased affinity for tRNAGln.

30 Glutaminyl-tRNA synthetase generates Gln-tRNA(Gln) 10(7)-fold more efficiently than Glu-tRNA(Gln)

31 zes major identity elements clustered in the tRNA(Gln) acceptor stem.

32 In addition, pyroglutamyl-tRNA(Gln) accumulated during the reaction catalyzed by t

33 However, Glu-tRNA(Gln) activates the glutaminase activity of the enzy

34 ional unit of the Bacillus subtilis glutamyl-tRNAGln amidotransferase have been cloned.

35 Gln-tRNAGln in B. subtilis and that glutamyl-tRNAGln amidotransferase is a novel and essential compon

36 ted evolution of Gln-tRNA synthetase and Glu-tRNAGln amidotransferase, and a novel, class I Lys-tRNA

37 by the glutamine-dependent Asp-tRNA(Asn)/Glu-tRNA(Gln) amidotransferase (Asp/Glu-Adt).

38 ) through the transamidation activity of Glu-tRNA(Gln) amidotransferase (Glu-AdT).

39 tRNA synthetase in a quaternary complex with tRNA(Gln), an ATP analog and glutamate reveals that the

40 yme is essential for the biosynthesis of Gln-tRNAGln, an obligate intermediate in translation.

41 inyl-tRNA synthetase (GlnRS) in complex with tRNAGln and ATP has identified a number a sequence-speci

42 dimensional structure of the complex between tRNAGln and glutaminyl-tRNA synthetase shows that the en

43 glutamine were cocrystallized with wild-type tRNAGln and their structures determined.

44 g glutamyl-tRNA synthetase to synthesize Glu-tRNA(Gln) and a glutaminyl-tRNA amidotransferase to conv

45 Many bacteria form Gln-tRNA(Gln) and Asn-tRNA(Asn) by conversion of the misacyl

46 GatCAB enzyme required in vivo for both Gln-tRNA(Gln) and Asn-tRNA(Asn) synthesis.

47 The amide aminoacyl-tRNAs, Gln-tRNA(Gln) and Asn-tRNA(Asn), are formed in many bacteria

48 NA(Asn) by conversion of the misacylated Glu-tRNA(Gln) and Asp-tRNA(Asn) species catalyzed by the Gat

49 Glutaminyl-tRNA(Gln) and asparaginyl-tRNA(Asn) were likely formed i

50 midation of the mischarged species, glutamyl-tRNA(Gln) and aspartyl-tRNA(Asn), by tRNA-dependent amid

51 s very low in the absence or presence of Glu-tRNA(Gln) and Gln.

52 king the NTD has reduced complementarity for tRNA(Gln) and glutamine.

53 NA(Gln) 10(7)-fold more efficiently than Glu-tRNA(Gln) and requires tRNA to synthesize the activated

54 ibit higher fidelity of 5'-maturation of pre-tRNA(Gln) and some of its mutant derivatives.

55 estor, used transamidation to synthesize Gln-tRNA(Gln) and that both the Bacteria and the Archaea ret

56 aminase but only in the presence of both Glu-tRNA(Gln) and the other subunit, GatE.

57 Eukaryotic tRNA(Gln) and tRNA(Glu) recognition determinants are fou

58 A synthetase (GluRS) that aminoacylates both tRNA(Gln) and tRNA(Glu) with glutamate.

59 their specific interactions with both A76 of tRNA(Gln)++ and glutamine.

60 ydrolysis, ATP hydrolysis, activation of Glu-tRNA(Gln), and aminolysis of activated tRNA by Gln-deriv

61 se (GluRS) first attaches glutamate (Glu) to tRNA(Gln), and an amidotransferase converts Glu-tRNA(Gln

62 ression by Cys-tRNA(Pro), Ser-tRNA(Thr), Glu-tRNA(Gln), and Asp-tRNA(Asn).

63 rmation of mis-acylated Asp-tRNA(Asn) or Glu-tRNA(Gln), and the subsequent amidation of these amino a

64 cts with nucleotides in the acceptor stem of tRNAGln, and at R260 in the enzyme's active site were fo

65 uncoupling of the first (1.72) base pair of tRNAGln, and tRNAMet was proposed by others to have a si

66 RS also separately differentiated to exclude tRNA(Gln) as a substrate, and the resulting discriminati

67 tion), Gln-competitive inhibition of the Glu-tRNA(Gln)/ATP-independent glutaminase activity of Glu-Ad

68 r transient kinetics experiments showed that tRNA(Gln) binds to GlnRS approximately 60-fold weaker wh

69 f an indirect aminoacylation pathway for Gln-tRNA(Gln) biosynthesis in Plasmodium that we hypothesize

70 These data show that Gln-tRNA(Gln) biosynthesis in the Plasmodium apicoplast proc

71 A is not formed by direct glutaminylation of tRNAGln but by a specific transamidation of Glu-tRNAGln.

72 The fact that only Glu-tRNA(Gln) but not tRNA(Gln) could activate the glutamina

73 tion of the weak first (U1-A72) base pair in tRNAGln by stacking between A72 and G2.

74 hia coli GlnRS to synthesize misacylated Glu-tRNA(Gln) by 16,000-fold.

75 Glu-tRNA(Gln), followed by conversion to Gln-tRNA(Gln) by a tRNA-dependent amidotransferase.

76 zymes found in organisms that synthesize Gln-tRNA(Gln) by an alternative pathway.

77 it was believed that most Bacteria form Gln-tRNA(GLN) by the amidation of Glu-tRNA(GLN), only a few

78 amidation of mischarged Asp-tRNA(Asn) or Glu-tRNA(Gln) catalyzed by a heterotrimeric amidotransferase

79 cation at U(34) of tRNA(Lys), tRNA(Glu), and tRNA(Gln), caused by the combination of eliminating the

80 fication at U34 of tRNA(Lys), tRNA(Glu), and tRNA(Gln) causes ribosome pausing at the respective codo

81 the RNA component of the contemporary GlnRS-tRNA(Gln) complex in mediating amino acid specificity.

82 structures of unliganded GlnRS and the GlnRS-tRNA(Gln) complex reveal that the Glu34 and Glu73 side c

83 NAPhe and tRNAAsp in the free state, and for tRNAGln complexed with glutaminyl-tRNA synthetase (GlnRS

84 st adopt the hairpinned conformation seen in tRNA(Gln) complexed with its synthetase.

85 uces Gln4 variants with reduced affinity for tRNA(Gln), consistent with a hinge-closing mechanism pro

86 duced by two to tenfold compared with native tRNA(Gln), consistent with previous findings that the te

87 The fact that only Glu-tRNA(Gln) but not tRNA(Gln) could activate the glutaminase activity of Gat

88 The construct is a modification of tRNAGln(CUA) from Tetrahymena thermophila, which natural

89 ne tRNAs: tRNA(Gln)(UUG), tRNA(Gln)(UUA) and tRNA(Gln)(CUA).

90 s were identified, defining positions in the tRNA(Gln)(CUG) anticodon stem that restrict first base w

91 Northern blot analysis revealed the sup70-65 tRNA(Gln)(CUG) is unstable, inefficiently charged, and 8

92 aturation of tRNA(Trp)(CCA), tRNA(Ile)(UAU), tRNA(Gln)(CUG), tRNA(Lys)(UUU), and tRNA(Val)(CAC).

93 mpromised expression of CAG-rich ORFs in the tRNA(Gln)(CUG)-depleted sup70-65 mutant.

94 iae, the SUP70 gene encodes the CAG-decoding tRNA(Gln)(CUG).

95 located in the anticodon and acceptor end of tRNAGln described previously.

96 t glutamylates both apicoplast tRNA(Glu) and tRNA(Gln), determined its kinetic parameters, and demons

97 y of specific interactions between GlnRS and tRNAGln determines amino acid affinity.

98 ing 9, 21, and 59 in tRNA(Cys) with those in tRNA(Gln) did not construct a functional core that conta

99 ision of Proteobacteria being able to charge tRNA(GLN) directly.

100 noacylate tRNA(Glu)versus those specific for tRNA(Gln) emerged and was experimentally validated.

101 that specific interactions between GlnRS and tRNAGln ensure the accurate positioning of the 3' termin

102 s primarily synthesized by first forming Glu-tRNA(Gln), followed by conversion to Gln-tRNA(Gln) by a

103 e (GlnRS) enzyme, which pairs glutamine with tRNA(Gln) for protein synthesis, evolved by gene duplica

104 transamidation pathway operates only for Gin-tRNAGln formation in this organism, and possibly in all

105 Here, we show a similar complex for Gln-tRNA(Gln) formation in Methanothermobacter thermautotrop

106 2 nM) sequesters the tRNA synthetase for Gln-tRNA(Gln) formation, with GatDE reducing the affinity of

107 GatCAB can be similarly used for Gln-tRNA(Gln) formation.

108 ormation, is not stable through product (Gln-tRNA(Gln)) formation, and has no major effect on the kin

109 icus GatCAB is capable of transamidating Glu-tRNA(Gln) from H. pylori or B. subtilis, and M. thermaut

110 isms lacking Gln-tRNA synthetase produce Gln-tRNA(Gln) from misacylated Glu-tRNA(Gln) through the tra

111 hrough the transamidation of misacylated Glu-tRNAGln, functionally replacing the lack of glutaminyl-t

112 ary core region of this species contains the tRNA(Gln) G15-C48 pair.

113 A specific allele of the SUP70/CDC65 tRNAGln gene (sup70-65) has been reported to be defectiv

114 zing amino acid activation suggests that the tRNAGln-GlnRS complex may be partly analogous to ribonuc

115 result different from that reported for the tRNAGln-glutaminyl-tRNA synthetase complex.

116 , and has no major effect on the kinetics of tRNA(Gln) glutamylation nor transamidation.

117 herichia coli glutaminyl-tRNA synthetase and tRNA(Gln) have been shown to determine the apparent affi

118 is of the crystal structure of tRNA(Cys) and tRNA(Gln) implicated long-range tertiary base-pairs abov

119 rthermore reconstituted in vitro cytoplasmic tRNAGln import into mitochondria by a novel mechanism.

120 tRNA(Gln) import into mammalian mitochondria proceeds by

121 noacylation determinants of Escherichia coli tRNAGln in a genetic and biochemical analysis of suppres

122 at transamidation is the only pathway to Gln-tRNAGln in B. subtilis and that glutamyl-tRNAGln amidotr

123 e showed in vivo localization of cytoplasmic tRNAGln in mitochondria and demonstrated its role in mit

124 using the latter product to transamidate Glu-tRNA(Gln) in concert with ATP hydrolysis.

125 ations at U(34) of tRNA(Lys), tRNA(Glu), and tRNA(Gln) in maintenance of mitochondrial genome, mitoch

126 Gln-tRNA(Gln) is synthesized from Glu-tRNA(Gln) in most microorganisms by a tRNA-dependent ami

127 ne tRNAs including tRNA(Lys), tRNA(Glu), and tRNA(Gln) in mto2/mss1, mto2/mto1, and mto2/mto1/mss1 st

128 Reexamination of the identity set of tRNA(Gln) in the light of these results indicates that i

129 ombinant GatAB converts Glu-tRNA(Gln) to Gln-tRNA(Gln) in vitro.

130 Archaea make glutaminyl-tRNA (Gln-tRNA(Gln)) in a two-step process; a non-discriminating g

131 Measurements of Gln-tRNA(Gln) interactions at the ribosome A-site show that

132 otransferase converted Asp-tRNA(Asn) and Glu-tRNA(Gln) into Asn-tRNA and Gln-tRNA, respectively.

133 ify Asp-tRNA(Asn) into Asn-tRNA(Asn) and Glu-tRNA(Gln) into Gln-tRNA(Gln); (iv) the TonB receptors an

134 Saccharomyces cerevisiae imports cytoplasmic tRNA(Gln) into the mitochondrion without any added prote

135 The Km of the T. thermophila enzyme for pre-tRNAGln is 1.6 x 10(-7)M, which is comparable to the val

136 Instead, tRNA(Gln) is initially acylated with glutamate by glutam

137 ryotic enzyme, whereas in other kingdoms Gln-tRNA(Gln) is primarily synthesized by first forming Glu-

138 aminyl-tRNA synthetase (GlnRS); instead, Gln-tRNA(Gln) is produced via an indirect pathway: a glutamy

139 Gln-tRNA(Gln) is synthesized from Glu-tRNA(Gln) in most micr

140 ey feature that distinguishes tRNA(Glu) from tRNA(Gln) is the third position in the anticodon of each

141 production of Gln-tRNA(Gln): misacylated Glu-tRNA(Gln) is transamidated by a Gln-dependent amidotrans

142 c hydrogen bond with U35 in the anticodon of tRNAGln, is involved in initial RNA recognition and is a

143 GluRS is active toward tRNA(Glu) and the two tRNA(Gln) isoacceptors the organism encodes, but with a

144 nto Asn-tRNA(Asn) and Glu-tRNA(Gln) into Gln-tRNA(Gln); (iv) the TonB receptors and ferric siderophor

145 he intermediate indicates that GatE is a Glu-tRNA(Gln) kinase.

146 inyl-tRNA synthetase (GlnRS) in complex with tRNAGln, leucine 136 (Leu136) stabilizes the disruption

147 three activities waning in concert when Glu-tRNA(Gln) levels become exhausted.

148 autotrophicus that allows the mischarged Glu-tRNA(Gln) made by the tRNA synthetase to be channeled to

149 alternate pathway for the production of Gln-tRNA(Gln): misacylated Glu-tRNA(Gln) is transamidated by

150 ered hybrid (GlnRS S1/L1/L2) synthesizes Glu-tRNA(Gln) more than 10(4)-fold more efficiently than Gln

151 l interdependence, crystal structures of two tRNA(Gln) mutants containing G15-G48 were determined bou

152 otransferase to convert Glu-tRNA(Gln) to Gln-tRNA(Gln) needed for protein synthesis.

153 hat GluRS2's primary role is to generate Glu-tRNAGln, not Glu-tRNAGlu.

154 a form Gln-tRNA(GLN) by the amidation of Glu-tRNA(GLN), only a few members of the gamma subdivision o

155 termediate to form the cognate products, Gln-tRNA(Gln) or Asn-tRNA(Asn).

156 midation of the mischarged tRNA species, Glu-tRNA(Gln) or Asp-tRNA(Asn).

157 ion of the mischarged tRNA species, glutamyl-tRNA(Gln) or aspartyl-tRNA(Asn).

158 as being able to form Asn-tRNA(Asn) and Gln-tRNA(Gln), our data demonstrate that while the enzyme is

159 E. coli tRNA(Gln) possesses the canonical Pu15-Py48 trans pairin

160 s jannaschii RPR's cis cleavage of precursor tRNA(Gln) (pre-tRNA(Gln)), which lacks certain consensus

161 onents formed cross-link products to the pre-tRNAGln probe.

162 otides at the acceptor and anticodon ends of tRNA(Gln) produced a tRNA substrate which was efficientl

163 vity by ATP or ATP-gammaS, together with Glu-tRNA(Gln), results either from an allosteric effect due

164 the class I glutaminyl-tRNA synthetase with tRNAGln revealed an uncoupling of the first (1.72) base

165 H. pylori contains one tRNAGlu and one tRNAGln species, whereas A. ferrooxidans possesses two o

166 We also show that both tRNA(Gln) species and a bacterial pre-tRNA(Asp) can be i

167 ndence of a G15-G48 Levitt pair, a number of tRNA(Gln) species containing G15-G48 were constructed an

168 ases that form the misacylated tRNA(Asn) and tRNA(Gln) species.

169 verting the M. thermautotrophicus GluRS to a tRNA(Gln) specific enzyme, solely through the addition o

170 upon to directly examine how GluRS2 acquired tRNA(Gln) specificity.

171 In contrast, Glu-tRNA(Gln) stimulates basal ATP hydrolysis slightly, but

172 GluRS to produce the required mischarged Glu-tRNAGln substrate.

173 modynamic framework for two-step cognate Gln-tRNA(Gln) synthesis demonstrates that the misacylating a

174 icus GluRS(ND), which is also capable of Glu-tRNA(Gln) synthesis, now shows that both k(cat) and K(m)

175 hetases to synthesize Asn and GatCAB for Gln-tRNA(Gln) synthesis, their AspRS enzymes were thought to

176 e phylogenetically diverse mechanisms of Gln-tRNA(Gln) synthesis.

177 introduction of G15-G48 into the non-cognate tRNA(Gln) tertiary core then significantly impairs CysRS

178 sence of GatDE has favored a unique archaeal tRNA(Gln) that may be preventing the acquisition of glut

179 rearranged, with the suite of genes encoding tRNA(Gln), the control region, and tRNA(Ile) located dow

180 In the absence of the amido acceptor, Glu-tRNA(Gln), the enzyme has basal glutaminase activity tha

181 by-product derived from gamma-phosphoryl-Glu-tRNA(Gln), the proposed high energy intermediate in Glu-

182 means for formation of correctly charged Gln-tRNAGln through the transamidation of misacylated Glu-tR

183 e produce Gln-tRNA(Gln) from misacylated Glu-tRNA(Gln) through the transamidation activity of Glu-tRN

184 In contrast, GluRS2 only misacylates tRNA(Gln) to form Glu-tRNA(Gln).

185 nstrated that recombinant GatAB converts Glu-tRNA(Gln) to Gln-tRNA(Gln) in vitro.

186 specialized amidotransferase to convert Glu-tRNA(Gln) to Gln-tRNA(Gln) needed for protein synthesis.

187 yze glutamine and were unable to convert Glu-tRNA(Gln) to Gln-tRNA(Gln) when glutamine was the amide

188 A(Gln), and an amidotransferase converts Glu-tRNA(Gln) to Gln-tRNA(Gln).

189 taminyl-tRNA amidotransferase to convert Glu-tRNA(Gln) to Gln-tRNA(Gln).

190 elongation factor binding to the cognate Gln-tRNA(Gln) together permit accurate protein synthesis wit

191 the proposed high energy intermediate in Glu-tRNA(Gln) transamidation.

192 three major glutamine tRNAs: tRNA(Gln)(UUG), tRNA(Gln)(UUA) and tRNA(Gln)(CUA).

193 , when marked versions of tRNA(Gln)(UUG) and tRNA(Gln)(UUA) flanked by identical sequences are expres

194 mJ target for Am in tRNA(Pro(GGG)) and Um in tRNA(Gln(UUG)) by mass spectrometric analysis.

195 A(Trp(CCA)) are substrates for Cm formation, tRNA(Gln(UUG)), tRNA(Pro(UGG)), tRNA(Pro(CGG)) and tRNA(

196 Finally, when marked versions of tRNA(Gln)(UUG) and tRNA(Gln)(UUA) flanked by identical s

197 mitochondrial import; thus sequences within tRNA(Gln)(UUG) direct import.

198 ear genes, and because ectopically expressed tRNA(Gln)(UUG) fractionates with mitochondria like its e

199 uclear and mitochondrial genetic codes, only tRNA(Gln)(UUG) has the capacity to function in mitochond

200 Because tRNA(Gln)(UUG) is a constituent of mitochondrial RNA fra

201 imately 10-20% of the cellular complement of tRNA(Gln)(UUG) is present in mitochondrial RNA fractions

202 complemented by overexpressing CAA-decoding tRNA(Gln)(UUG), an inefficient wobble-decoder of CAG.

203 thermophila has three major glutamine tRNAs: tRNA(Gln)(UUG), tRNA(Gln)(UUA) and tRNA(Gln)(CUA).

204 The activation of Glu-tRNA(Gln) via gamma-phosphorylation bears a similarity t

205 tRNA(Gln) was mainly cytosolic in localization; tRNA(Ile

206 Here, guided by the core of E. coli tRNA(Gln), we sought to test and identify alternative fu

207 Neither tRNA(Glu) nor tRNA(Gln) were substrates.

208 were unable to convert Glu-tRNA(Gln) to Gln-tRNA(Gln) when glutamine was the amide donor.

209 R's cis cleavage of precursor tRNA(Gln) (pre-tRNA(Gln)), which lacks certain consensus structures/seq

210 However, the core of E. coli tRNA(Gln), which contains G15:C48, is functional for cys

211 how GluRS2 achieves specific recognition of tRNA(Gln) while rejecting the two H. pylori tRNA(Glu) is

212 lutamyl-tRNA synthetase (ND-GluRS) forms Glu-tRNA(Gln), while the heterodimeric amidotransferase GatD

213 e enzyme transamidates Asp-tRNA(Asn) and Glu-tRNA(Gln) with similar efficiency (k(cat)/K(m) of 1368.4