Aminoacyl-transfer RNA populations in mammalian cells. Chromatographic profiles and patterns of codon recognition

Hatfield, Dolph L.; Matthews, C. Robert; Rice, Marlen C.

doi:10.1016/0005-2787(79)90032-7

Cited by 75 publications

(54 citation statements)

References 36 publications

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“…The aminoacylation of tRNA [Ser]Sec proceeded slowly and reached a maximal level at Ϸ4 min, and then the seryl-tRNA [Ser]Sec product began to deacylate. The rapid decline in product was most certainly due to the amount of ATP used in reaction mixtures, which was much lower than that previously used in generating seryl-tRNA [Ser]Sec with mammalian seryltRNA synthetases (14). Phosphorylation of seryl-tRNA [Ser]Sec occurred rapidly and reached a plateau in Ϸ3-4 min (Fig.…”

Section: Resultsmentioning

confidence: 90%

“…1 Ser was used as a substrate for PSTK in 50 l of reaction mixture [20 mM Tris⅐HCl (pH 7.4)͞0.01 mM EGTA͞1 mM DTT͞10 mM MgCl 2 ͞50 M ATP͞1 l of [␥-32 P]ATP (final specific activity Ϸ600 Ci͞mmol)͞0.5 g of pure PSTK]. Coupled aminoacylation-phosphorylation reactions involved the same components plus rabbit reticulocyte aminoacyl-tRNA synthetases (14). Reactions were incubated for 20 min or at time intervals designated in figures at 30°C and were spotted on P81 phosphocellulose filter paper.…”

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confidence: 99%

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Identification and characterization of phosphoseryl-tRNA ^[Ser]Sec kinase

Carlson

Kryukov

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

174

187

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In 1970, a kinase activity that phosphorylated a minor species of seryl-tRNA to form phosphoseryl-tRNA was found in rooster liver [Maenpaa, P. H. & Bernfield, M. R. (1970) Proc. Natl. Acad. Sci. USA 67, 688 -695], and a minor seryl-tRNA that decoded the nonsense UGA was detected in bovine liver. The phosphoseryl-tRNA and the minor UGA-decoding seryl-tRNA were subsequently identified as selenocysteine (Sec) tRNA [Ser]Sec , but the kinase activity remained elusive. Herein, by using a comparative genomics approach that searched completely sequenced archaeal genomes for a kinase-like protein with a pattern of occurrence similar to that of components of Sec insertion machinery, we detected a candidate gene for mammalian phosphoseryl-tRNA [ S elenocysteine (Sec) has its own code word, UGA, and its own tRNA, and therefore is viewed as the 21st amino acid in the genetic code (reviewed in refs. 1-4). Although UGA usually codes for the termination of protein synthesis, it also specifies Sec if specific requirements are met. The presence of a stemloop structure downstream of UGA, called a Sec insertion sequence (SECIS) element, is the critical component in selenoprotein mRNAs that dictates UGA to code for Sec (reviewed in ref. 5). In mammals, the SECIS element occurs in the 3Ј untranslated region of selenoprotein mRNAs. A specific elongation factor, EFsec, specifically recognizes selenocysteyltRNA [Ser]Sec (6, 7), and a SECIS element binding protein, SBP2, binds specifically to the SECIS element (8), directing the insertion of Sec into protein in response to UGA.It has been known for several years that the biosynthesis of Sec occurs on its tRNA in both bacteria (9) and mammals (10) after the tRNA is initially aminoacylated with serine by seryl-tRNA synthetase. In Escherichia coli, the pathway for the biosynthesis of Sec has been completely established (reviewed in ref. 1). After the aminoacylation of bacterial tRNA [Ser]Sec with serine, the hydroxyl group is removed from the seryl moiety to yield an aminoacrylyl intermediate, and this step is catalyzed by a pyridoxal phosphate-dependent Sec synthase. The aminoacrylyl intermediate serves as the acceptor for the activated selenium donor, monoselenophosphate, which is synthesized from selenite and ATP in the presence of selenophosphate synthetase (reviewed in ref. 1). Once selenium is donated to the intermediate, the biosynthesis of Sec on tRNA [Ser]Sec is complete.In eukaryotes, however, the biosynthesis of Sec has not been established, but several components have been identified over the years that play a role in this process. For example, in 1970, a minor seryl-tRNA was reported to form phosphoseryl-tRNA by a kinase activity from rooster liver (11), and a minor seryl-tRNA from bovine, rabbit, and chicken livers was reported to specifically decode the nonsense codon UGA (12). Although it was subsequently shown that both the minor seryl-tRNA that formed phosphoseryl-tRNA and the one that decoded UGA were the same tRNA (13), it was not known at the time these components were ...

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Section: Resultsmentioning

confidence: 90%

mentioning

confidence: 99%

Identification and characterization of phosphoseryl-tRNA ^[Ser]Sec kinase

Carlson

Kryukov

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

174

187

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“…tRNA was prepied from rabbit liver and [3H]Lys-tRNA prepared using rabbit reticulocyte synthetases [6]. A portion of the [3H]Lys-tRNA was resolved over a RPCd column [7], the resulting [3H]Lys-tRN& isolated, resolved a second time over the RPC-5 column, isolated and added to a total i3H] Lys-tRNA preparation in order to enrich for ['HJLys-tRN&.…”

Section: Methodsmentioning

confidence: 99%

Relative utilization of mammalian Lys‐tRNA isoacceptors in protein synthesis

et al. 1980

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“…Total RNA was extracted from 300 grams (wet weight) of yeast cells grown in YPD medium (Rubin 1975) and tRNA was purified by DEAE-cellulose chromatography and deacylated (Hatfield et al 1979), all as previously described. Transfer RNAs were aminoacylated with [ 3 H]methionine (70 Ci/mmole, Amersham) or [ 35 S]methionine (1000 Ci/mmole; Amersham) under conditions of limiting tRNA and the labeled tRNAs were fractionated by reversed-phase chromatography on a RPC-5 column (Kelmers and Heatherly 1971) essentially as described (Hatfield et al 1979), except that elution of aminoacyl-tRNAs was carried out using a linear gradient of 0.45-0.65 M NaCl in the presence of 10 mM magnesium acetate.…”

Section: Analysis Of Trna Modificationmentioning

confidence: 99%

“…Transfer RNAs were aminoacylated with [ 3 H]methionine (70 Ci/mmole, Amersham) or [ 35 S]methionine (1000 Ci/mmole; Amersham) under conditions of limiting tRNA and the labeled tRNAs were fractionated by reversed-phase chromatography on a RPC-5 column (Kelmers and Heatherly 1971) essentially as described (Hatfield et al 1979), except that elution of aminoacyl-tRNAs was carried out using a linear gradient of 0.45-0.65 M NaCl in the presence of 10 mM magnesium acetate. For HPLC analysis of base modifications, the tRNA was digested to nucleosides by nuclease P1 and alkaline phosphatase (Gehrke et al 1982) and the hydrolyzate was analyzed by HPLC according to the method of Gehrke and Kuo (1990).…”

Section: Analysis Of Trna Modificationmentioning

confidence: 99%

The essential Gcd10p–Gcd14p nuclear complex is required for 1-methyladenosine modification and maturation of initiator methionyl-tRNA

Anderson

Phan²,

Cuesta³

et al. 1998

Genes Dev.

238

327

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Gcd10p andMet maturation. The chromatographic behavior of elongator and initiator tRNA Met on a RPC-5 column indicated that both species are altered structurally in gcd10⌬ cells, and analysis of base modifications revealed that 1-methyladenosine (m 1 A) is undetectable in gcd10⌬ tRNA. Interestingly, gcd10 and gcd14 mutations had no effect on processing or accumulation of elongator tRNA Met , which also contains m 1 A at position 58, suggesting a unique requirement for this base modification in initiator maturation.

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Aminoacyl-transfer RNA populations in mammalian cells. Chromatographic profiles and patterns of codon recognition

Cited by 75 publications

References 36 publications

Identification and characterization of phosphoseryl-tRNA ^[Ser]Sec kinase

Identification and characterization of phosphoseryl-tRNA ^[Ser]Sec kinase

Relative utilization of mammalian Lys‐tRNA isoacceptors in protein synthesis

The essential Gcd10p–Gcd14p nuclear complex is required for 1-methyladenosine modification and maturation of initiator methionyl-tRNA

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Aminoacyl-transfer RNA populations in mammalian cells. Chromatographic profiles and patterns of codon recognition

Cited by 75 publications

References 36 publications

Identification and characterization of phosphoseryl-tRNA [Ser]Sec kinase

Identification and characterization of phosphoseryl-tRNA [Ser]Sec kinase

Relative utilization of mammalian Lys‐tRNA isoacceptors in protein synthesis

The essential Gcd10p–Gcd14p nuclear complex is required for 1-methyladenosine modification and maturation of initiator methionyl-tRNA

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Identification and characterization of phosphoseryl-tRNA ^[Ser]Sec kinase

Identification and characterization of phosphoseryl-tRNA ^[Ser]Sec kinase