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 efficiently and specifically hydrolyzes Ala-tRNAPro.

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

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

5 iency prevents the removal of the downstream tRNAPro.

6 vate cysteine and to mischarge cysteine onto tRNAPro.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

32 hermobacter thermautotrophicus that enhances tRNA(Pro) aminoacylation.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

73 2 was confirmed as the TrmJ target for Am in tRNA(Pro(GGG)) and Um in tRNA(Gln(UUG)) by mass spectrom

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

118 Whereas all tRNAPro sequences from bacteria strictly conserve A73 an

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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