ライフサイエンスコーパス: tRNA(Pro)

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

19 Ala-tRNAPro is specifically hydrolyzed by the editing domain

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

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

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

23 efficiently and specifically hydrolyzes Ala-tRNAPro.

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

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

26 Whereas all tRNAPro sequences from bacteria strictly conserve A73 an

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

66 iency prevents the removal of the downstream tRNAPro.

67 hermobacter thermautotrophicus that enhances tRNA(Pro) aminoacylation.

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

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

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

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

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

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

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

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

76 2 was confirmed as the TrmJ target for Am in tRNA(Pro(GGG)) and Um in tRNA(Gln(UUG)) by mass spectrom

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

107 vate cysteine and to mischarge cysteine onto tRNAPro.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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