ライフサイエンスコーパス: tRNA(Pro

1 ed to decrease the levels of proline-charged tRNA(Pro) .

2 must be present in the peptidyl site, e.g., tRNA(Pro).

3 TTC-3' sequence found in the T psi C loop of tRNA(Pro).

4 nthesize both cysteinyl-tRNA(Cys) and prolyl-tRNA(Pro).

5 the noncognate amino acid before transfer to tRNA(Pro).

6 yptophan, a peptidyl-tRNA also appears, TnaC-tRNA(Pro).

7 or ATP and proline, but not proline alone or tRNA(Pro).

8 tion of MA membrane binding than full-length tRNA(Pro).

9 y to confer the full inhibitory effects upon tRNA(Pro).

10 nt tRNA methylation site in S. pombe, C34 of tRNA(Pro).

11 like protein, is responsible for editing Ala-tRNA(Pro).

12 in (INS) but lack the capability to edit Cys-tRNA(Pro).

13 (ProRSs) mischarge alanine and cysteine onto tRNA(Pro).

14 lyzes smaller Ala-tRNA(Pro) and excludes Pro-tRNA(Pro).

15 ackbone interactions in recognition of human tRNA(Pro).

16 eukemia virus (MuLV) preferentially captures tRNA(Pro).

17 meric enzyme, with specificity for yeast Ala-tRNA(Pro).

18 or its nonfunctional substitute, TnaC(W12R)-tRNA(Pro).

19 the putative site occupied by Trp12 of TnaC-tRNA(Pro).

20 vate cysteine and to mischarge cysteine onto tRNAPro.

21 domain, is capable of weakly deacylating Ala-tRNAPro.

22 efficiently and specifically hydrolyzes Ala-tRNAPro.

23 a-microhelixPro variants but not cognate Pro-tRNAPro.

24 to be complementary to the 3' 18 nt of human tRNAPro.

25 iency prevents the removal of the downstream tRNAPro.

26 y complex formation between ProRS, YbaK, and tRNAPro.

27 Previous work had shown that the tRNA(Pro) acceptor stem elements A73 and G72 (both stric

28 showed that base-specific recognition of the tRNA(Pro) acceptor stem is critical for recognition by E

29 ng experiments confirmed that the end of the tRNA(Pro) acceptor stem is proximal to this motif 2 loop

30 ursor containing both RNase P RNA (RPM1) and tRNA(Pro) accumulated dramatically.

31 For Methanococcus jannaschii tRNA(Pro), accuracy is difficult because the cognate pro

32 so abolished the protection of U2609 by TnaC-tRNA(Pro) against chemical methylation.

33 , the removal of the m(1)G37 modification of tRNA(Pro) also disrupts U32-A38 pairing in a structurall

34 ated aminoacyl-adenylate, a prerequisite for tRNA(Pro) aminoacylation.

35 hermobacter thermautotrophicus that enhances tRNA(Pro) aminoacylation.

36 c INS domain, was capable of deacylating Ala-tRNAPro and Ala-microhelixPro variants but not cognate P

37 K binds to ProRS to gain specificity for Cys-tRNAPro and avoid deacylation of Cys-tRNACys in the cell

38 of a three-component complex with ProRS and tRNAPro and establish the stoichiometry of a 'triple-sie

39 YbaK and show that it efficiently edits Cys-tRNAPro and that a conserved Lys residue is essential fo

40 t editing domains that function to clear Ala-tRNA(Pro) and Cys-tRNA(Pro) in vivo.

41 lished the specific attachment of proline to tRNA(Pro) and cysteine to tRNA(Cys).

42 s" studies at these two positions of E. coli tRNA(Pro) and determined that major groove functional gr

43 tRNA synthetases that hydrolyzes smaller Ala-tRNA(Pro) and excludes Pro-tRNA(Pro).

44 in part, by elements in the acceptor stem of tRNA(Pro) and further ensured through collaboration with

45 ine structure to discriminate against prolyl-tRNA(Pro) and promote termination in the absence of a st

46 codon or acceptor stem, the two hotspots for tRNA(Pro) and tRNA(Cys) identity determinants.

47 ssess the dual capacity to aminoacylate both tRNA(Pro) and tRNA(Cys) with their cognate amino acids.

48 escentus ProRS can readily form Cys- and Ala-tRNA(Pro), and deacylation studies confirmed that these

49 acid was efficiently acylated in vitro onto tRNA(Pro), and the misacylated Cys-tRNA(Pro) was not edi

50 ch as CC[C/U]-[C/U], read by isoacceptors of tRNA(Pro), are highly prone to +1 frameshift (+1FS) erro

51 uctures of the bacterial ribosome containing tRNA(Pro) bound to either cognate or slippery codons to

52 roRS) have been shown to misacylate Cys onto tRNA(Pro), but lack a Cys-specific editing function.

53 when RNAs that contain the anticodon arm of tRNA(Pro), but not that of tRNA(Lys3), are added exogeno

54 The deacylation rate of Ala-tRNA(Pro) by At ProXp-ala was also significantly reduced

55 here that, in some respects, recognition of tRNA(Pro) by M. jannaschii ProRS parallels that of human

56 es that are inhibited are hydrolysis of TnaC-tRNA(Pro) by release factor 2 and peptidyl transfer of T

57 s critical for the hydrolytic editing of Ala-tRNA(Pro) by this class II synthetase.

58 The absence of m(1)G37 in tRNA(Pro) causes +1 frameshifting on polynucleotide, sli

59 m formation, tRNA(Gln(UUG)), tRNA(Pro(UGG)), tRNA(Pro(CGG)) and tRNA(His(GUG)) for Um, and tRNA(Pro(G

60 etermination of the steady-state kinetics of tRNA(Pro) charging showed that the catalytic efficiency

61 ee tryptophan binding and inhibition of TnaC-tRNA(Pro) cleavage at the peptidyl transferase center.

62 phan prevents sparsomycin inhibition of TnaC-tRNA(Pro) cleavage with wild-type ribosome complexes but

63 ibosome where bound tryptophan inhibits TnaC-tRNA(Pro) cleavage.

64 The ribosome-TnaC-tRNA(Pro) complexes analyzed were formed in vitro; they

65 nucleotide A2572 of wild-type ribosome-TnaC-tRNA(Pro) complexes but not of ribosome-TnaC(W12R)-tRNA(

66 ro) complexes but not of ribosome-TnaC(W12R)-tRNA(Pro) complexes.

67 An analysis of chimeric tRNA(Pro) constructs showed that, in addition to A73 and

68 aminoacylation by human ProRS on a chimeric tRNAPro containing the E. coli acceptor-TpsiC stem-loop

69 A73 and G72, transplantation of the E. coli tRNA(Pro) D-domain was necessary and sufficient to conve

70 a novel substrate-assisted mechanism of Cys-tRNA(Pro) deacylation that prevents nonspecific Pro-tRNA

71 by tryptophan is primarily a consequence of tRNA(Pro) depletion, resulting from TnaC-tRNA(Pro) reten

72 diting domain that deacylates mischarged Ala-tRNAPro, editing of Cys-tRNAPro has not been demonstrate

73 Sequence differences in the tRNA-proline (tRNApro) end of the mitochondrial control-region of thre

74 A missense mutant strain, which requires Cys-tRNA(Pro) for growth in the absence of thymine.

75 phan was not as efficient in protecting TnaC-tRNA(Pro) from puromycin action as wild-type ribosomes.

76 nscribed with its substrates, tRNA met f and tRNAPro, from a promoter located upstream of the tRNA me

77 product catalyzes the m(1)G37 methylation of tRNA(Pro) Furthermore, substitution of three of the four

78 located 5' to the mt tRNA(fMet)-RNase P RNA-tRNA(Pro) gene cluster, so that the mitochondrially enco

79 N), tRNASer(AGN), tRNAMet(AUA), tRNATrp, and tRNAPro genes occur in M. californianus mitochondria, st

80 One consists of 27 tRNA(Pro) genes and the other contains 27 tandem repeats

81 f the tRNA genes have introns, including the tRNAPro (GGG) gene, which contains a second intron at an

82 2 was confirmed as the TrmJ target for Am in tRNA(Pro(GGG)) and Um in tRNA(Gln(UUG)) by mass spectrom

83 RNA(Pro(CGG)) and tRNA(His(GUG)) for Um, and tRNA(Pro(GGG)) for Am. tRNA(Ser(UGA)), previously observ

84 lates mischarged Ala-tRNAPro, editing of Cys-tRNAPro has not been demonstrated and a double-sieve mec

85 kpoints to prevent formation of Ala- and Cys-tRNA(Pro) have been described, including the Ala-specifi

86 residues leads to a significant loss in Ala-tRNA(Pro) hydrolysis, and altering the size of the pocke

87 o) deacylation that prevents nonspecific Pro-tRNA(Pro) hydrolysis.

88 was reduced by the presence of Trp12 in TnaC-tRNA(Pro), implying A788 displacement.

89 we show that the imino acid proline and not tRNAPro imposes the primary eIF5A requirement for polypr

90 g wherein a third sieve is used to clear Cys-tRNAPro in at least some organisms.

91 , a free-standing editing domain, clears Cys-tRNAPro in trans.

92 la is another editing domain that clears Ala-tRNAPro in trans.

93 e-domain INS homolog, YbaK, which clears Cys-tRNA(Pro) in trans.

94 recently shown to hydrolyze misacylated Cys-tRNA(Pro) in trans.

95 that function to clear Ala-tRNA(Pro) and Cys-tRNA(Pro) in vivo.

96 idyl-tRNA of the tna operon of E. coli, TnaC-tRNA(Pro), in the presence of excess tryptophan, resists

97 and a fast mechanism during translocation of tRNA(Pro) into the P-site.

98 Additionally, E. coli tRNAPro is a poor substrate for human ProRS, and the pre

99 esis that the specificity of YbaK toward Cys-tRNAPro is determined by the formation of a three-compon

100 Ala-tRNAPro is specifically hydrolyzed by the editing domain

101 In contrast, Cys-tRNA(Pro) is cleared by a freestanding INS domain homolo

102 Instead, Cys-tRNA(Pro) is cleared by a single-domain homolog of INS,

103 Mischarged Ala-tRNA(Pro) is hydrolyzed by an editing domain (INS) prese

104 t the unmodified transcript of M. jannaschii tRNA(Pro) is indeed mis-acylated with cysteine.

105 These findings establish that Trp-12 of TnaC-tRNA(Pro) is required for introducing specific changes i

106 cur by two mechanisms, a slow mechanism when tRNA(Pro) is stalled in the P-site next to an empty A-si

107 ommodation, is rate limiting for natural Pro-tRNA(Pro) isoacceptors.

108 ptophan inhibited puromycin cleavage of TnaC-tRNA(Pro); it also inhibited binding of the antibiotic s

109 due at position 12 of the peptidyl-tRNA TnaC-tRNA(Pro) leads to the creation of a free tryptophan bin

110 Further evolution of the Af-tRNA(Pro) led to a variant exhibiting significantly impr

111 se effects were not observed with TnaC(W12R)-tRNA(Pro) mutant complexes.

112 Nevertheless, studies with different tRNA(Pro) mutants in Salmonella enterica suggest that fr

113 S errors requires the m(1)G37 methylation of tRNA(Pro) on the 3' side of the anticodon and the transl

114 vitro; they contained either wild-type TnaC-tRNA(Pro) or its nonfunctional substitute, TnaC(W12R)-tR

115 mes rephased on this sequence, with peptidyl-tRNA(Pro) pairing with CCC in the +1 frame.

116 e ribosome, and the role of the nascent TnaC-tRNA(Pro) peptide in facilitating tryptophan binding and

117 coli, interactions between the nascent TnaC-tRNA(Pro) peptidyl-tRNA and the translating ribosome cre

118 The presence of the TnaC-tRNA(Pro) peptidyl-tRNA within the ribosome protects the

119 no acid sequence of TnaC of the nascent TnaC-tRNA(Pro) peptidyl-tRNA, in addition to the presence of

120 r of the eukaryotic-like group, although its tRNA(Pro) possesses prokaryotic features in the acceptor

121 conditions the accumulation of Arg(12)-TnaC-tRNA(Pro) prevented Rho-dependent transcription terminat

122 substrates, specificities for removal of the tRNAPro primer and polypurine tract stability are lost,

123 ex consists of the last four residues of the tRNA(Pro) primer for (-) strand DNA synthesis of Moloney

124 le structured regions in both the U5-PBS and tRNA(Pro) primer that otherwise sequester residues neces

125 of unspliced and spliced viral RNA, and the tRNA(Pro) primer was properly annealed to the primer bin

126 ommodated into the ribosome and bound to Pro-tRNA(Pro), productive synthesis of the peptide bond occu

127 naschii ProRS catalyzes the synthesis of Cys-tRNA(Pro) readily, the enzyme is unable to edit this mis

128 ease in the inhibitory effect relative to WT tRNA(Pro), replacing the entire D arm sequence with that

129 nM and 45 nM in the absence and presence of tRNA(Pro), respectively.

130 on of the D loop sequence of tRNA(Lys3) into tRNA(Pro) resulted in a modest increase in the inhibitor

131 of tRNA(Pro) depletion, resulting from TnaC-tRNA(Pro) retention within stalled, translating ribosome

132 Our structures of NC bound to U5-PBS and tRNA(Pro) reveal the structure-based mechanism for retro

133 Whereas all tRNAPro sequences from bacteria strictly conserve A73 an

134 C1.G72, all available cytoplasmic eukaryotic tRNAPro sequences have a C73 and a G1.C72 base pair.

135 n the context of missense suppression by Cys-tRNA(Pro), Ser-tRNA(Thr), Glu-tRNA(Gln), and Asp-tRNA(As

136 o be sufficient to eliminate all misacylated tRNAPro species from the cell.

137 the aminoacyl-ester bond of misacylated Ala-tRNA(Pro) species.

138 g translation by hydrolyzing misacylated Ala-tRNA(Pro) that has been synthesized by prolyl-tRNA synth

139 elieved by overexpression of tRNA(1)(Pro), a tRNA(Pro) that translates CCG, but not CCU.

140 acid analogues using an optimized suppressor tRNA(Pro) that we designed.

141 dons mediate the response to proline-charged tRNA(Pro), the levels of which decrease under proline li

142 me that has just completed synthesis of TnaC-tRNA(Pro), the peptidyl-tRNA precursor of the leader pep

143 sents the uORF2 peptide covalently linked to tRNA(Pro), the tRNA predicted to decode the carboxy-term

144 was determined by crosslinking Lys11 of TnaC-tRNA(Pro) to nucleotide A750 of 23S rRNA.

145 ctor 2 and peptidyl transfer of TnaC of TnaC-tRNA(Pro) to puromycin.

146 ication revealed that MuLV prefers to select tRNA(Pro), tRNA(Gly), or tRNA(Arg).

147 substrates for Cm formation, tRNA(Gln(UUG)), tRNA(Pro(UGG)), tRNA(Pro(CGG)) and tRNA(His(GUG)) for Um

148 ine-adenylate (Ala-AMP) and a mischarged Ala-tRNAPro variant, respectively.

149 diting, and (3) deacylating a mischarged Ala-tRNA(Pro) variant via a post-transfer editing pathway.

150 able of rapidly deacylating a mischarged Ala-tRNA(Pro) variant.

151 s depleted of release factor 2, Arg(12)-TnaC-tRNA(Pro) was accumulated in the absence or presence of

152 itro onto tRNA(Pro), and the misacylated Cys-tRNA(Pro) was not edited by ProRS.

153 Using misacylated Pro-tRNAPhe and Phe-tRNAPro, we show that the imino acid proline and not tRN

154 d tRNA(Leu), the mitochondrial tRNA(Val) and tRNA(Pro)) were strongly associated with the observed ra

155 toward a particular anticodon variant of Af-tRNA(Pro), whereas others are promiscuous.

156 t, the INS domain is unable to deacylate Cys-tRNA(Pro), which is hydrolyzed exclusively by a freestan

157 Human tRNA(Pro), which lacks these elements, is not aminoacyla

158 ry, in which we altered the PBS to anneal to tRNA(Pro), while simultaneously randomizing the viral RN

159 ) is a potent inhibitor of aminoacylation of tRNAPro with broad biomedical applications.

160 Prolyl-tRNA synthetases (ProRS) mischarge tRNA(Pro) with alanine or cysteine due to their smaller

161 showing that M. jannaschii ProRS misacylates tRNA(Pro) with cysteine, and argue against the proposal

162 naschii ProRS misaminoacylates M. jannaschii tRNA(Pro) with cysteine.

163 lack of m(1)G37 destabilize interactions of tRNA(Pro) with the P site of the ribosome, causing large

164 o how m(1)G37 stabilizes the interactions of tRNA(Pro) with the ribosome in the context of a slippery

165 olyl-tRNA synthetases are known to mischarge tRNA(Pro) with the smaller amino acid alanine and with c

166 yptophan inhibits puromycin cleavage of TnaC-tRNA(Pro) with wild-type ribosome complexes, it does not

167 ered Archaeoglobus fulgidus prolyl-tRNAs (Af-tRNA(Pro)) with three different anticodons: CUA, AGGG, a