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 efficiently rejected than the fully modified tRNA(Lys).

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

8 nism by repairing 5'-PO4 and 3'-OH groups in 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 ndary structure of the HIV genomic RNA-human tRNA(Lys)(3) initiation complex using heteronuclear nucl

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

47 cleotides comprising the anticodon domain of tRNALys,3.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

83 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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

131 76 patients with VAP for integration at this tRNA(lys) recombination site demonstrated that patients

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

158 17 nucleotide anticodon stem-loop of E. coli tRNA(Lys) was then assembled from these synthons using p

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

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

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

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

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

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

165 ing mechanism which prevents misacylation of tRNALys with ornithine.

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

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