ライフサイエンスコーパス: tRNA(Lys

1 fluorescently labeled substrate (BODIPY-Lys-tRNA(Lys)).

2 and catalyses the specific aminoacylation of tRNA(Lys).

3 omes to the level of native Escherichia coli tRNA(Lys).

4 RS, and GagPol interacting with both Gag and tRNA(Lys).

5 ic for Ala-tRNA(Ala), and MprF2 utilizes Lys-tRNA(Lys).

6 nism by repairing 5'-PO4 and 3'-OH groups in tRNA(Lys).

7 efficiently rejected than the fully modified tRNA(Lys).

8 nslocation is slower with the s(2)-deficient tRNA(Lys).

9 no and Roth and previously thought to affect tRNA(Lys).

10 ulture: one contained a PBS complementary to tRNA(Lys)1,2, while the second maintained a PBS compleme

11 The human tRNA(Lys)3 gene was used as a target for transposition i

12 mentary to the 3'-terminal 18 nucleotides of tRNA(Lys)3, we identified an HIV-1 virus which contained

13 rted back to the wild type, complementary to tRNA(Lys)3.

14 We have developed a mutant tRNA(Lys)(3) derivative, tRNA(Lys)(3)A58U, in which A58

15 of the 18-bp primer helix with the 3' end of tRNA(Lys)(3) drives large conformational rearrangements

16 id dissociation kinetics of RT from the vRNA-tRNA(Lys)(3) initiation complex and reveal a specific st

17 ndary structure of the HIV genomic RNA-human tRNA(Lys)(3) initiation complex using heteronuclear nucl

18 tRNA(Lys)(3) interacts directly with HIV-1 reverse trans

19 rse transcriptase (RT) initiates from a host tRNA(Lys)(3) primer bound to the vRNA genome and is the

20 e present study, we investigated the role of tRNA(Lys)(3) residue A58 in the replication cycle of HIV

21 Cellular tRNA(Lys)(3) serves as the primer for reverse transcript

22 complex formed between a host transfer RNA (tRNA(Lys)(3)) and a region at the 5' end of genomic RNA;

23 the viral genomic RNA or the cognate primer, tRNA(Lys)(3), was apparently unaffected.

24 Residue A58 of tRNA(Lys)(3), which lies outside the PBS-complementary r

25 developed a mutant tRNA(Lys)(3) derivative, tRNA(Lys)(3)A58U, in which A58 was replaced by U.

26 e catalytic site, a two piece semi-synthetic tRNA(Lys, 3)construct was used.

27 ty between nucleotides at the 5' terminus of tRNA(Lys,3) and the U5-IR loop of the feline immunodefic

28 pe 1 (HIV-1), the 3' 18 nucleotides of human tRNA(Lys,3) are annealed to a complementary sequence on

29 HIV-1 reverse transcriptase uses human tRNA(Lys,3) as a primer to initiate reverse transcriptio

30 In contrast, correctly sized tRNA(Lys,3) can be prepared by (i) total chemical synthe

31 he zinc finger structures, is able to anneal tRNA(Lys,3) efficiently to the PBS, and to destabilize t

32 aining full-length or 5'-deleted variants of tRNA(Lys,3) hybridized to the primer-binding site.

33 Human tRNA(Lys,3) is the specific primer for HIV-1 reverse tra

34 modified bases on the efficiency with which tRNA(Lys,3) is used in vitro as the HIV-1 replication pr

35 imer-binding site with natural and synthetic tRNA(Lys,3) primers, indicating it was not a consequence

36 gated the effects of the corresponding human tRNA(Lys,3) versions of the E. coli modifications, using

37 n human immunodeficiency virus type 1, human tRNA(Lys,3), is selectively packaged into the virion alo

38 The human analogue, tRNA(Lys,3), is the specific tRNA primer for HIV-1 rever

39 he anticodon stem-loop (ASL) domain of human tRNA(Lys,3), the primer for HIV-1 reverse transcriptase.

40 of anticodon positions U35 and U36 of human tRNA(Lys,3).

41 when primed with either natural or synthetic tRNA(Lys,3).

42 In human tRNA(Lys,3)UUU three modified bases are present in the a

43 ild-type binding activity of wild-type human tRNA(Lys,3)UUU.

44 the acceptor-TPsiC stem-loop domain of human tRNA(Lys,3)was not specifically aminoacylated by the hum

45 These undermodified tRNALys,3 anticodon loops are distinctly different from

46 anticodon recognition and for utilization of tRNALys,3 by HIV-1 as the native reverse transcriptase p

47 Viral Pol proteins influenced tRNAlys,3 packaging but had little influence on virion p

48 ication and the additional A+-C base-pair on tRNALys,3 structure.

49 cleotides comprising the anticodon domain of tRNALys,3.

50 t from those for viral genomic RNA or primer tRNAlys,3.

51 All mutants showed reduced affinity for tRNALys-3 and supported significantly less (-)-strand DN

52 sis that mutant PBS reversion is a result of tRNALys-3 annealing onto and extension from a PBS that s

53 of HIV-1 RT supports an interaction with the tRNALys-3 anticodon loop critical for efficient (-)-stra

54 V type 1 (HIV-1) specifically uses host cell tRNALys-3 as a primer for reverse transcription.

55 Structural features of binary RT.tRNALys-3 complexes were examined by in situ footprintin

56 We have developed a mutant tRNALys-3 derivative with mutations in the PBS-binding r

57 tRNALys-3 interacts directly with HIV-1 reverse transcri

58 rast, NC promotes specific annealing of only tRNALys-3 onto an RNA template (HXB2) whose PBS sequence

59 ements outside the acceptor-TPsiC domains of tRNALys-3 play an important role in preferential primer

60 Cellular tRNALys-3 serves as the primer for reverse transcription

61 Recently, tRNALys-3 was cross-linked via its anticodon loop to hum

62 uences quickly revert to be complementary to tRNALys-3.

63 idosis and stroke-like episodes (MELAS); the tRNA(Lys) 8344 mutation causing myoclonic epilepsy and r

64 The intergenic COII/tRNA(Lys) 9-bp deletion in human mtDNA, which is found a

65 in homoplasmic form either the mitochondrial tRNA(Lys) A8344G mutation associated with the myoclonic

66 as that the G2.U71 wobble pair of spirochete tRNALys acts as antideterminant for class II LysRS but d

67 , we show how the s(2) modification in yeast tRNA(Lys) affects mRNA decoding and tRNA-mRNA translocat

68 otype caused by the m.8344A>G mutation in mt-tRNA(Lys), aminoacylated by a Class II aaRS.

69 EF-G-catalyzed translocation step of the two tRNALys and the slippery codons from the A- and P- sites

70 e unfolding process of charged and uncharged tRNALys and tRNALeu(UUR) has revealed that the separatio

71 The altered tRNA(Lys) and luciferase genes were introduced into Nico

72 le for 2-thiouridylation of mt-tRNA(Glu), mt-tRNA(Lys) and mt-tRNA(Gln).

73 pe and mutant human mt-tRNA(Leu(UUR)) and mt-tRNA(Lys), and stabilize mutant mt-tRNA(Leu(UUR)).

74 gested that the yjdF aptamer is a homolog of tRNA(Lys), and that two of the conserved loops of the ri

75 Purified Arg-tRNALys, Thr-tRNALys, and Met-tRNALys were essentially not deacylated

76 f structure-based sequence alignments, seven tRNALys anticodon variants and seven LysRS1 anticodon bi

77 their ability to recognize Escherichia coli tRNA(Lys) anticodon mutants.

78 s of unmodified and pseudouridine39-modified tRNA(Lys) anticodon stem loops (ASLs) show that signific

79 for the fully modified 17-nucleotide E. coli tRNA(Lys) anticodon stem-loop domain (ASL).

80 d UUUAAAG (40%) (underlined codon decoded by tRNA(Lys), anticodon 5' mnm5s2UUU 3') was more complex,

81 in the anticodon domain of Escherichia coli tRNA(Lys) are necessary for high-affinity codon recognit

82 harging pyrrolysine to tRNA(Pyl); lysine and tRNA(Lys) are not substrates of the enzyme.

83 t the anticodon stem loops for tRNA(Glu) and tRNA(Lys) are substrates of comparable activity to the f

84 lass I LysRSs recognize the same elements in tRNALys as their class II counterparts, namely the discr

85 as seen in the recent crystal structures of tRNA(Lys) ASLs bound to the 30S ribosomal subunit.

86 m(5)s(2)U34, s(2)U34, t(6)A37, and Mg(2+) on tRNA(Lys) ASLs to decipher how the E. coli modifications

87 ne), or at positions -2, -6 and -10 (for the tRNA(Lys)AUC gene), and repression reached 90%.

88 sequences encoding the Arabidopsis thaliana tRNA(Lys)AUC or tRNA(Trp)AUC suppressor tRNAs, and tRNA

89 cific recognition of the same nucleotides in tRNALys by the two unrelated types of enzyme suggests th

90 rrangements in the ribosome-EF-Tu-GDP-Pi-Lys-tRNA(Lys) complex following GTP hydrolysis by EF-Tu.

91 The anticodon domain of E. coli tRNA(Lys) contains the hypermodified nucleosides mnm(5)s

92 uggest that specificity for the anticodon of tRNALys could have been acquired through relatively few

93 sphatidylglycerol synthase (A-PGS) or by Lys-tRNA(Lys)-dependent lysyl-phosphatidylglycerol synthase

94 ng to both the cytoplasmic and mitochondrial tRNA(Lys), despite the difference in the discriminator b

95 ture of the analogous t6A containing E. coli tRNA(Lys), despite the presence of the bulky methylthio

96 Refinement of the LysRS1-tRNALys docking model based upon these data suggested th

97 analysis of the Pyrococcus horikoshii LysRS1-tRNALys docking model.

98 dition of a G between positions 36 and 37 of tRNA(Lys) expand the anticodons of both tRNAs similarly

99 nzyme whose major function is to provide Lys-tRNALys for protein synthesis, also catalyzes aminoacyla

100 rtions have been identified in the T loop of tRNALys from Didymium and tRNAGlu from Physarum.

101 of nucleotide (nt) 8344 in the mitochondrial tRNALys gene, were examined for the proportion of mutant

102 egion containing avrPphB was inserted into a tRNALYS gene, which was re-formed at the right junction

103 A nuclear tRNA(Lys) gene from Arabidopsis thaliana was cloned and

104 The plasmid subsequently integrated into a tRNA(Lys) gene in the chromosome of each recipient, wher

105 Cell lines carrying the MERRF mitochondrial tRNA(Lys) gene mutation, which causes a pronounced decre

106 A novel G8363A mutation in the mtDNA tRNA(Lys) gene was associated, in two unrelated families

107 polymorphic region of the chromosome near a tRNA(Lys) gene, suggesting that exoU is a horizontally a

108 tegration site within the 3' terminus of the tRNA(Lys) gene.

109 API-1 can reintegrate into either of the two tRNA(Lys) genes, including the one that was used for int

110 er, PAPI-1 integrates into either of the two tRNA(Lys) genes.

111 Mutation of tRNA(Ala) and tRNA(Lys) had little effect on either MprF activity, ind

112 Native tRNA(Lys) has a U-turn structure similar to the yeast tR

113 (hLysRS) is essential for aminoacylation of tRNA(Lys) Higher eukaryotic LysRSs possess an N-terminal

114 However, decreased lysyl-tRNA(Lys) in the lysS mutant provides a possible rationa

115 stal structures of the ribosome containing a tRNA(Lys) in the P site with a U*U mismatch with the mRN

116 he large ribosomal subunit and a Cy5-labeled tRNA(Lys) in the ribosomal peptidyl-tRNA-binding (P) sit

117 This observation indicates that the tRNALys initiation step plays an important role in the d

118 ng length variation analysis of the COII and tRNALYS intergenic region, nucleotide sequence analysis

119 sRS), are required for specific packaging of tRNALys into virions.

120 ) was more complex, since the wobble base of tRNA(Lys) is modified at two positions.

121 intergenic region of cytochrome c oxidase II/tRNA(Lys), is the most common mitochondrial deletion.

122 that suggest the importance of the ratio of tRNALys isoacceptors in Type-2 diabetes.

123 nown to interact specifically with all three tRNA(Lys) isoacceptors, is also selectively packaged int

124 RNA(Lys) packaging complex that includes the tRNA(Lys) isoacceptors, LysRS, HIV-1 Gag, GagPol, and vi

125 inetic analysis we show that mcm(5)-modified tRNA(Lys) lacking the s(2) group has a lower affinity of

126 ng genomic islands to the corresponding PAO1 tRNA(Lys)-linked genomic island, the pathogenicity islan

127 aging, a Gag.GagPol complex interacts with a tRNA(Lys).LysRS complex, with Gag interacting specifical

128 The Escherichia coli tRNA(Lys) modifications mnm(5)s(2)U34 and t(6)A37 have i

129 ex (MSC), restricting the pool of free LysRS-tRNA(Lys) Mounting evidence suggests that LysRS is relea

130 cherichia coli unable to modify fully either tRNA(Lys) or tRNA(Asn).

131 ne)-encoding mRNAs conducted with either Lys-tRNA(Lys) or Val-tRNA(Lys) reveals that that amino acid

132 d that LeuRS specifically reduced the Km for tRNA(Lys) over 3-fold, with no additional change seen up

133 We have previously defined a tRNA(Lys) packaging complex that includes the tRNA(Lys)

134 s studies support the hypothesis that during tRNA(Lys) packaging, a Gag.GagPol complex interacts with

135 two unrelated types of enzyme suggests that tRNALys predates at least one of the LysRSs in the evolu

136 inary complex does not occur when a chimeric tRNALys/Pro containing proline-specific D and anticodon

137 decoding AAG in the ribosomal A-site, E.coli tRNA(Lys) promotes a highly unusual single-tRNA slippage

138 76 patients with VAP for integration at this tRNA(lys) recombination site demonstrated that patients

139 s conducted with either Lys-tRNA(Lys) or Val-tRNA(Lys) reveals that that amino acid charge, while imp

140 tRNA(Lys) sampling and accommodation to the empty A site

141 To examine the specificity of tRNALys selection, E. coli tRNA3Lys was modified to tRNA

142 l synthetase/tRNA pair derived from archaeal tRNA(Lys) sequences that efficiently and selectively inc

143 ass I-type enzyme to aminoacylate particular tRNALys species and provides a molecular basis for the o

144 fic anticodon domain modified nucleosides of tRNA(Lys) species would restore ribosomal binding and al

145 s while expression of the amber and missense tRNA(Lys) suppressor genes from a geminivirus vector cap

146 The slipperiness of tRNA(Lys), therefore, cannot be ascribed to a single mod

147 Purified Arg-tRNALys, Thr-tRNALys, and Met-tRNALys were essentially n

148 to be a methylthiotransferase that modifies tRNA(Lys) to enhance translational fidelity of transcrip

149 reviously that one class of MprF can use Lys-tRNA(Lys) to modify phosphatidylglycerol (PG), but the m

150 While the delivery of BOP-Lys-tRNA(Lys) to the ribosome is limited by its poor binding

151 at the wobble position of the anticodons of tRNA(Lys), tRNA(Glu), and tRNA(1)(Gln).

152 of the mcm(5) or s(2) modification at U34 of tRNA(Lys), tRNA(Glu), and tRNA(Gln) causes ribosome paus

153 a critical role of modifications at U(34) of tRNA(Lys), tRNA(Glu), and tRNA(Gln) in maintenance of mi

154 no detectable levels of nine tRNAs including tRNA(Lys), tRNA(Glu), and tRNA(Gln) in mto2/mss1, mto2/m

155 ompletely abolished modification at U(34) of tRNA(Lys), tRNA(Glu), and tRNA(Gln), caused by the combi

156 nomethyl (cmnm)(5)s(2)U(34) in mitochondrial tRNA(Lys), tRNA(Glu), and tRNA(Gln).

157 et of tRNAs, including tRNA(Glu), tRNA(Gly), tRNA(Lys), tRNA(Val), tRNA(His), tRNA(Asp), and tRNA(SeC

158 recognize the third anticodon nucleotide of tRNA(Lys) (U36) and those that recognize both the second

159 ng effects of tRNA overexpression, implicate tRNA(Lys(UUU)) as a target of EcoPrrC toxicity in yeast.

160 was selectively rescued by overexpression of tRNA(Lys) UUU as well by overexpression of genes (BCK1 a

161 Elevated tRNA(Lys)UUU levels suppressed the elp3Delta phenotypes

162 ph3Delta phenotypes, indicating that lack of tRNA(Lys)UUU modifications were responsible.

163 tRNA structure in the fundamentally dynamic tRNA(Lys)(UUU) anticodon.

164 The data suggest that tRNA(Lys)(UUU) species require anticodon domain modifica

165 hat catalyzes methylthiolation of t(6)A37 in tRNA(Lys)(UUU) to ms(2)t(6)A37.

166 s-linked reads originating from AAA-decoding tRNA(Lys)(UUU) were 10-fold enriched over its cellular a

167 A(Trp)(CCA), tRNA(Ile)(UAU), tRNA(Gln)(CUG), tRNA(Lys)(UUU), and tRNA(Val)(CAC).

168 17 nucleotide anticodon stem-loop of E. coli tRNA(Lys) was then assembled from these synthons using p

169 transfer (smFRET) between (Cy5)EF-G and (Cy3)tRNALys, we studied the translational elongation dynamic

170 Purified Arg-tRNALys, Thr-tRNALys, and Met-tRNALys were essentially not deacylated by LysRS, indica

171 nly cytosolic in localization; tRNA(Ile) and tRNA(Lys) were mainly mitochondrial; and tRNA(Trp) and t

172 uring coded protein synthesis requires lysyl-tRNA(Lys), which is synthesized by lysyl-tRNA synthetase

173 synthesis, also catalyzes aminoacylation of tRNALys with arginine, threonine, methionine, leucine, a

174 trand contains the UUU anticodon sequence of tRNALys with flanking GCs to increase duplex stability.

175 ing mechanism which prevents misacylation of tRNALys with ornithine.

176 e show that MnmA binds to unmodified E. coli tRNA(Lys) with affinity in the low micromolar range.

177 RS, including the propensity to aminoacylate tRNA(Lys) with suboptimal identity elements, as well as