Structural requirements of tRNALys for its import into yeast mitochondria

被引:56
作者
Entelis, NS
Kieffer, S
Kolesnikova, OA
Martin, RP
Tarassov, IA
机构
[1] Univ Strasbourg 1, Inst Physiol & Chim Biol, CNRS,Div Cellulaire & Dev, Unite Propre Rech 9005, F-67084 Strasbourg, France
[2] Moscow MV Lomonosov State Univ, Fac Biol, Dept Mol Biol, Moscow 119899, Russia
关键词
D O I
10.1073/pnas.95.6.2838
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
In the yeast Saccharomyces cerevisiae, one of the two cytoplasmic lysine tRNAs, tRNA(CUU)(Lys), is partially associated with the mitochondrial matrix. Mitochondrial import of this tRNA requires binding to the precursor of the mitochondrial lysyl-tRNA synthetase, pre-MSK, and aminoacylation by the cytoplasmic lysyl-tRNA synthetase, KRS, appears to he a prerequisite for this binding. The second lysine isoacceptor tRNA(mnm5s2UUU)(LYS) {where 5-[(methylamino)-methyl]-2-thiouridine is mnm(5)s(2)U} is exclusively localized in the cytoplasm. To study import determinants within the tRNA(CUU)(Lys) molecule, we introduced a panel of replacements in the original sequences of the imported and nonimported lysine tRNAs that correspond to domains or individual residues that differ between these two isoacceptors. The mutant transcripts were tested for import, aminoacylation, and binding to pre-MSK, Import and aminoacylation efficiencies correlate well for the majority of mutant transcripts. However, some poorly aminoacylated transcripts were rather efficiently imported. Surprisingly, these transcripts retained binding capacity to pre-MSK. In fact, all imported transcripts retained pre-MSK binding capacity but nonimported versions did not, suggesting that this binding, rather than aminoacylation, is essential for import. Substitution of the anticodon arm of tRNA(CUU)(Lys), with that of tRNA(mnm5s2UUU)(Lys) abolished import without affecting aminoacylation. A version of tRNA(mnm5s2UUU)(Lys) with an anticodon CUU was efficiently imported in vitro and was also found to be imported in vivo. This implies that the anticodon arm, especially position 34, is important for recognition by the import machinery. A nicked tRNA(CUU)(Lys) transcript is still imported but its import requires reannealing of the two tRNA moieties, which implies that tRNA(CUU)(Lys) is imported as a folded molecule.
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页码:2838 / 2843
页数:6
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共 37 条
[1]  
CIRAKOGLU B, 1985, EUR J BIOCHEM, V149, P353
[2]  
DELREY FJ, 1982, P NATL ACAD SCI-BIOL, V79, P4138
[3]   Editing and import: Strategies for providing plant mitochondria with a complete set of functional transfer RNAs [J].
Dietrich, A ;
Small, I ;
Cosset, A ;
Weil, JH ;
MarechalDrouard, L .
BIOCHIMIE, 1996, 78 (06) :518-529
[4]   FACTORS DETERMINING SPECIFICITY OF TERTIARY RNA AMINOACYLATION REACTION - NON-ABSOLUTE SPECIFICITY OF TERTIARY RNA-AMINOACYL TERTIARY RNA-SYNTHETASE RECOGNITION AND PARTICULAR IMPORTANCE OF MAXIMAL VELOCITY [J].
EBEL, JP ;
GIEGE, R ;
BONNET, J ;
KERN, D ;
BEFORT, N ;
BOLLACK, C ;
FASIOLO, F ;
GANGLOFF, J ;
DIRHEIMER, G .
BIOCHIMIE, 1973, 55 (05) :547-557
[5]   Mitochondrial import of a yeast cytoplasmic tRNA(Lys): Possible roles of aminoacylation and modified nucleosides in subcellular partitioning [J].
Entelis, NS ;
Krasheninnikov, IA ;
Martin, RP ;
Tarassov, IA .
FEBS LETTERS, 1996, 384 (01) :38-42
[6]   ANTICODON-INDEPENDENT AMINOACYLATION OF AN RNA MINIHELIX WITH VALINE [J].
FRUGIER, M ;
FLORENTZ, C ;
GIEGE, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (09) :3990-3994
[7]   STRUCTURE AND EVOLUTION OF A GROUP OF RELATED AMINOACYL-TRANSFER RNA-SYNTHETASES [J].
GATTI, DL ;
TZAGOLOFF, A .
JOURNAL OF MOLECULAR BIOLOGY, 1991, 218 (03) :557-568
[8]   Interplay of tRNA-like structures from plant viral RNAs with partners of the translation and replication machineries [J].
Giege, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (22) :12078-12081
[9]   TRANSFER-RNA STRUCTURE AND AMINOACYLATION EFFICIENCY [J].
GIEGE, R ;
PUGLISI, JD ;
FLORENTZ, C .
PROGRESS IN NUCLEIC ACID RESEARCH AND MOLECULAR BIOLOGY, VOL 45, 1993, 45 :129-206
[10]   TRANSFER-RNAS ARE IMPORTED INTO MITOCHONDRIA OF TRYPANOSOMA-BRUCEI INDEPENDENTLY OF THEIR GENOMIC CONTEXT AND GENETIC-ORIGIN [J].
HAUSER, R ;
SCHNEIDER, A .
EMBO JOURNAL, 1995, 14 (17) :4212-4220