Studies on T. Utilis tRNATYr variants with enzymatically altered D-loop sequences. II. relationship between the tertiary structure and tyrosine acceptance

Takashi Ohyama, Kazuya Nishikawa, Shosuke Takemura

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The nucleotide sequence of T. utilis tRNATYr has been modified to have a deletion or substitution of the "conserved" nucleotide sequence Gm18-G19 in the D-loop by enzymatic procedures in vitro. Conformations of the variant tRNAs were analyzed by measuring melting profiles and electrophoretic mobiities in "native" polyacrylamide gels, and by examining the RNase T1 digestion patterns in sequencing gels. The results obtained shed light on the importance of the interaction between the sequence Gm18-G19 and nucleotides in the TPSgr;C-loop (probably PSgr;57-C59 for the maintenance of the total conformation of tRNATYr in solution. The association of D-loop and TΨC-loop regions in the variant tRNATyrs is slightly relaxed even at room temperature and melting occurred at temperatures higher than 40°C. The relationship between the tertiary structure of the variant tRNA and its aminoacylation capacity was assayed at various temperatures. The results indicate that highly ordered tertiary structure is needed for tRNATyr to be fully aminoacylated.

Original languageEnglish
Pages (from-to)859-866
Number of pages8
JournalJournal of Biochemistry
Volume99
Issue number3
Publication statusPublished - 1986
Externally publishedYes

Fingerprint

Nucleotides
Tyrosine
Transfer RNA
Freezing
Temperature
Conformations
Melting
Gels
RNA, Transfer, Tyr
Transfer RNA Aminoacylation
Conformation
Ribonuclease T1
Conserved Sequence
Polyacrylates
Digestion
Substitution reactions
Maintenance
Association reactions
Deletion
Sequencing

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Physiology (medical)
  • Radiology Nuclear Medicine and imaging
  • Molecular Biology
  • Biochemistry

Cite this

Studies on T. Utilis tRNATYr variants with enzymatically altered D-loop sequences. II. relationship between the tertiary structure and tyrosine acceptance. / Ohyama, Takashi; Nishikawa, Kazuya; Takemura, Shosuke.

In: Journal of Biochemistry, Vol. 99, No. 3, 1986, p. 859-866.

Research output: Contribution to journalArticle

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N2 - The nucleotide sequence of T. utilis tRNATYr has been modified to have a deletion or substitution of the "conserved" nucleotide sequence Gm18-G19 in the D-loop by enzymatic procedures in vitro. Conformations of the variant tRNAs were analyzed by measuring melting profiles and electrophoretic mobiities in "native" polyacrylamide gels, and by examining the RNase T1 digestion patterns in sequencing gels. The results obtained shed light on the importance of the interaction between the sequence Gm18-G19 and nucleotides in the TPSgr;C-loop (probably PSgr;57-C59 for the maintenance of the total conformation of tRNATYr in solution. The association of D-loop and TΨC-loop regions in the variant tRNATyrs is slightly relaxed even at room temperature and melting occurred at temperatures higher than 40°C. The relationship between the tertiary structure of the variant tRNA and its aminoacylation capacity was assayed at various temperatures. The results indicate that highly ordered tertiary structure is needed for tRNATyr to be fully aminoacylated.

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