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2 ic rate constants for pre-tRNA(met608) and a pre-tRNA(met608) (+1)C/(+72)A mutant provides evidence t
5 gnize and cleave any target RNA that forms a pre-tRNA-like complex with another RNA, in some cases cl
6 he highly structured T stem-loop region in a pre-tRNA substrate is bound by the B. subtilis P RNA.
8 e S-domain binds the T stem-loop region in a pre-tRNA substrate to confer specificity for tRNA substr
9 get mRNA to form a tertiary structure like a pre-tRNA and recruit intracellular ribonuclease P (RNase
10 imately 4-fold better than the cleavage of a pre-tRNA substrate by the wild-type RNase P RNA under th
11 The 2'OH group at the cleavage site of a pre-tRNA substrate is an important determinant in the cl
12 tional groups in the T stem-loop region of a pre-tRNA substrate previously identified to directly con
14 ubstrate selectivity is investigated using a pre-tRNA substrate containing single-atom modifications
16 nzyme reaction is much more efficient with a pre-tRNA substrate, binding at least 100-fold better and
17 bonucleoprotein enzyme RNase P processes all pre-tRNAs, yet some substrates apparently lack consensus
18 trate that is cleaved slowly by P RNA alone (pre-tRNA(f-met605)) and one that is cleaved quickly (pre
20 re cleaved only after the discriminator, and pre-tRNA variants with a total of N bp (N is less than 1
23 uence-specific contact between P protein and pre-tRNA that contributes to molecular recognition of RN
24 cross-linked nucleotides in the ribozyme and pre-tRNA were determined by reverse transcriptase primer
30 a and dysfunctional processing of associated pre-tRNAs that are spliced but 5' and 3' unprocessed, wi
31 he ability to cleave the covalently attached pre-tRNA, indicating that the cross-links reflect the na
34 n a scenario elucidated by the use of the av pre-tRNAs, algal permuted tRNA genes could have further
35 ting of a small group I intron from Azoarcus pre-tRNA(Ile) showed that tertiary interactions between
37 that both tRNA(Gln) species and a bacterial pre-tRNA(Asp) can be imported in vitro into mitochondria
38 f1 via small interfering RNA increased basal pre-tRNA and rendered tRNA synthesis refractory to mTOR
40 These findings demonstrate a link between pre-tRNA splicing and pre-mRNA 3' end formation, suggest
43 Herein we describe the use of biotinylated pre-tRNA substrates to isolate RNase P ribozyme-substrat
46 r cytoplasmic recruitment, packaging of both pre-tRNAs and U6 snRNA requires the nuclear export recep
47 component of bacterial RNase P can catalyse pre-tRNA cleavage in the absence of the RNase P protein
48 ecause some early steps in the S. cerevisiae pre-tRNA biosynthetic pathway are nucleolar, we examined
52 ities and cleavage rates of Escherichia coli pre-tRNAs that exhibit the largest variation from consen
53 ly characterized substrate-enzyme conjugate [pre-tRNA(Tyr)-Methanocaldococcus jannaschii (Mja) RPR] t
55 (SEN) during processing of intron-containing pre-tRNAs and by Ire1 cleavage of HAC1 mRNA following in
57 studies of the location of intron-containing pre-tRNAs in the rna1-1 mutant rule out the possibility
59 rom postmitotic neurons results in defective pre-tRNA and pre-rRNA processing and progressive neurode
60 pathways for tRNA maturation, with defective pre-tRNAs being most sensitive to decay and most depende
61 d nuclear accumulation, prevented disordered pre-tRNA processing, and restored suppression, indicatin
65 cleavage of precursor tRNA in vivo, enhances pre-tRNA binding by directly contacting the 5'-leader se
66 hylation of the two nucleotides of exogenous pre-tRNA(Trp) added to an H. volcanii cell extract also
69 . subtilis RNase P has a higher affinity for pre-tRNA with adenosine at N(-4), and this binding prefe
71 nce of apparent catalytic rate constants for pre-tRNA(met608) and a pre-tRNA(met608) (+1)C/(+72)A mut
72 k(cat)/K(m) (by approximately 360-fold) for pre-tRNA cleavage to those observed with partially purif
76 equence-specific contacts between the fourth pre-tRNA nucleotide on the 5' side of the cleavage site
77 s, the enzyme catalyzing intron removal from pre-tRNA is a heterotetrameric complex (splicing endonuc
80 uclease (3' tRNase) removes 3' trailers from pre-tRNAs by cleaving the RNA immediately downstream of
81 s that perform the same biological function, pre-tRNA maturation, thereby providing insight into the
83 within polycistronic transcripts to generate pre-tRNAs that subsequently become substrates for RNase
84 i RPR's cis cleavage of precursor tRNA(Gln) (pre-tRNA(Gln)), which lacks certain consensus structures
87 Recent work has demonstrated that the host pre-tRNA processing enzyme, RNase P, can cleave the HCV
93 ct tRNA nuclear export also cause defects in pre-tRNA splicing leading to tight coupling of the splic
95 RNase P protein is to offset differences in pre-tRNA structure such that binding and catalysis are u
96 ests that some aspects of their functions in pre-tRNA and pre-rRNA processing pathways might overlap
97 tin-t homologue, binds tRNA and functions in pre-tRNA splicing and export of mature tRNA from the nuc
98 y conserved regions, the regions involved in pre-tRNA recognition and the location of the active site
101 portin for tRNA, suppresses the reduction in pre-tRNA levels, AAM gene up-regulation, and slow growth
105 , and J18/2 regions of ribozyme structure in pre-tRNA binding and implicate an additional region, J11
110 Considering that perturbed intranuclear pre-tRNA metabolism and apparent deficiency in tRNA nucl
111 ific C-terminal domain (CTD) of hLa maintain pre-tRNA in an unprocessed state by blocking the 5'-proc
113 es accumulation of nuclear and mitochondrial pre-tRNAs, suggesting that JhI-1 encodes both forms of t
116 leader sequence of precursor tRNA molecules (pre-tRNA), whereas the protein subunit assists in substr
117 Consistently, overexpression of a mutant pre-tRNA(Tyr) that cannot be processed by RNase P had a
119 t deficiency in the processing of the mutant pre-tRNA, that becomes limiting for protein synthesis on
120 s could affect tRNase Z processing of mutant pre-tRNAs, perhaps contributing to mitochondrial disease
125 experiments showed that formation of native pre-tRNA is delayed by misfolding of P3-P9, including mi
126 equences form separate domains in the native pre-tRNA, their folding is coupled via metastable non-na
134 and in bacteria the RNA alone is capable of pre-tRNA processing in vitro, i.e. it is a catalytic RNA
138 protein component alter the pH dependence of pre-tRNA(Asp) cleavage catalyzed by RNase P, providing f
140 of contact between the mature tRNA domain of pre-tRNA and the ribozyme; however, relatively little is
145 nonical editing events, within the intron of pre-tRNA(Tyr)GUA, involving guanosine-to-adenosine trans
146 These data demonstrate that the 5' leader of pre-tRNA moves 4 to 6 A closer to the PRNA x P protein i
147 irs with the 5' leader increases the K(m) of pre-tRNA slightly, in agreement with previous results.
149 ently been shown to facilitate maturation of pre-tRNA, and three distinct ribosomal proteins, Rpl16p,
151 re we investigate the enzymatic mechanism of pre-tRNA hydrolysis catalyzed by the NYN (Nedd4-BP1, Yac
152 bases encoding the +1 and +2 nucleotides of pre-tRNA Tyr, had a strong deleterious effect in vivo, a
153 zation is required for the normal pathway of pre-tRNA maturation, facilitates assembly of small RNAs
154 that operates upstream of a major pathway of pre-tRNA maturation, which itself is upstream of tRNA ex
156 0-fold decrease in the self-cleavage rate of pre-tRNA(Tyr)-MjaDeltaU RPR compared to the wild type, a
157 t in RNase P is to facilitate recognition of pre-tRNA by enhancing the interaction between the enzyme
158 c endonuclease that catalyzes the removal of pre-tRNA leader sequences to form the 5' end of mature t
159 cleavage by Escherichia coli RNase P RNA of pre-tRNA in which specific pro-Rp phosphate oxygens were
160 th the single-stranded 5' leader sequence of pre-tRNA, and (ii) the orientation and register of the p
161 photoagent was incorporated into a series of pre-tRNA substrates containing unique uridine residues i
162 The mutation causes the anticodon stem of pre-tRNA(Arg)(CCG) to misfold into an alternative helix
163 logy and demonstrate that all three steps of pre-tRNA splicing, as well as tRNA nuclear export and am
165 nuclease protection assays with a variety of pre-tRNA substrates and mutant La proteins indicate that
169 eal that a greater-than-expected fraction of pre-tRNAs from both E. coli and B. subtilis contains a n
171 pathway that monitors both end processing of pre-tRNAs and the modification state of mature tRNAs.
173 protein subunit and the leader sequences of pre-tRNAs may be common in bacterial RNase P and may lea
174 tes that bind and protect the 3' trailers of pre-tRNAs from exonuclease digestion via sequence-specif
178 for the wild-type pre-tRNA(Ser)CGA and other pre-tRNAs, Lhp1p is required for the normal endonucleoly
184 n-coding RNA, snoRNAs and, most prominently, pre-tRNAs and other Pol III transcripts are targeted for
186 (f-met605)) and one that is cleaved quickly (pre-tRNA(met608)) pinpoint the characteristic C(+1)/A(+7
187 endonuclease (TSEN) complex, largely reduced pre-tRNA cleavage activity, and accumulation of linear t
188 length and that this significantly restricts pre-tRNAs to a La-independent pathway of maturation in f
189 of the leader to 2 nucleotides due to P RNA-pre-tRNA contacts that are stabilized by the P protein.
191 RNA mutations: a mutation can impede several pre-tRNA processing steps, with each such reduction cont
193 ts for 3' tRNase substrates, we tested small pre-tRNA(Arg) substrates lacking the D and anticodon ste
196 We propose that binding by Lhp1p stabilizes pre-tRNAs in conformations that allow the 3' endonucleol
197 molecule which by itself can bind substrate pre-tRNA, select and hydrolyze the correct phosphodieste
198 ribonuclease P (RNase P) binds to substrate pre-tRNAs with high affinity and catalyzes site-specific
202 d guide nucleotides, we now demonstrate that pre-tRNA methylation is guided in trans by the intron-en
205 thylation-competent box C/D RNPs on both the pre-tRNA and the excised intron (both linear and circula
206 The 3'-end trailer can be removed by the pre-tRNA processing endonuclease tRNase Z, an ancient, c
207 activity is dependent on 3' oligo(U) in the pre-tRNA for interaction with the N-terminal RNA binding
208 the first position in the palindrome in the pre-tRNA sequence, which does not affect tRNA function.
210 s part of the catalytic pocket formed in the pre-tRNA-RNase P complex and participates in the binding
211 a1p corrects the nuclear export, but not the pre-tRNA-splicing defects of los1Kan(r) cells, thereby u
212 roteins to determine the conformation of the pre-tRNA 5' leader relative to the protein in the holoen
214 separate the 5' from the 3' terminus of the pre-tRNA and to position the cleavage site in the cataly
215 auses partial or complete degradation of the pre-tRNA by RNase R, whereas extension of the stem resul
216 and (ii) the orientation and register of the pre-tRNA leader sequence in the central cleft places the
217 cal data and three-dimensional models of the pre-tRNA showed that the tRNA is folded, and that the tR
219 so eliminate the obligatory refolding of the pre-tRNA that would be required to carry out two cis-met
220 ed region, likely alters the geometry of the pre-tRNA-binding cleft and may provide a functional expl
222 A uses a specificity module to recognize the pre-tRNA and a catalytic module to perform cleavage.
223 mase family, endonucleolytically removes the pre-tRNA 3' trailer in a step central to tRNA maturation
225 ribozyme that are in close proximity to the pre-tRNA cleavage site, we introduced the short-range ph
227 ow the conserved nucleotides adjacent to the pre-tRNA substrate contribute to substrate binding and p
228 ing that binding of these antibiotics to the pre-tRNA substrate contributes to the inhibitory activit
229 formations of the two 3 nt bulges, where the pre-tRNA is cleaved, are stabilized by stacking interact
230 for tgm silencing regardless of whether the pre-tRNA transcripts are substrates for Mod5 modificatio
231 Nucleoli were slightly fragmented, and the pre-tRNAs went from their normal, mostly nucleolar locat
232 box C/D guide RNA within the intron of this pre-tRNA led to the assumption that nucleotide methylati
234 ugh binding by the yeast La protein Lhp1p to pre-tRNAs is required for the normal pathway of tRNA mat
251 strong accumulation of all tested unspliced pre-tRNA species, as well as accumulation of 5' and 3' u
257 nism of 2'-O-methylation for the H. volcanii pre-tRNA(Trp) in vitro by assembling methylation-compete
261 urification-tagged Lsm3p was associated with pre-tRNA primary transcripts and, less efficiently, with
264 independently, interactions of RNase P with pre-tRNA(Tyr) containing either the 5' leader, the 3' tr
265 e formation of complexes of RNase P RNA with pre-tRNA or tRNA, and at least one site stabilizes the t
267 haromyces cerevisiae La homologue Lhp1p with pre-tRNAs was reduced approximately threefold on depleti
270 nces the magnesium affinity of the RNase P x pre-tRNA complex indirectly by binding and positioning p
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