A protein extension to shorten RNA: Elongated elongation factor-Tu recognizes the D-arm of T-armless tRNAs in nematode mitochondria

Masayuki Sakurai, Yoh Ichi Watanabe, Kimitsuna Watanabe, Takashi Ohtsuki

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Nematode mitochondria possess extremely truncated tRNAs. Of 22 tRNAs, 20 lack the entire T-arm. The T-arm is necessary for the binding of canonical tRNAs and EF (elongation factor)-Tu (thermo-unstable). The nematode mitochondrial translation system employs two different EF-Tu factors named EF-Tu1 and EF-Tu2. Our previous study showed that nematode Caenorhabditis elegans EF-Tu1 binds specifically to T-armless tRNA. C. elegans EF-Tu1 has a 57-amino acid C-terminal extension that is absent from canonical EF-Tu, and the T-arm-binding residues of canonical EF-Tu are not conserved. In this study, the recognition mechanism of T-armless tRNA by EF-Tu1 was investigated. Both modification interference assays and primer extension analysis of cross-linked ternary complexes revealed that EF-Tu1 interacts not only with the tRNA acceptor stem but also with the D-arm. This is the first example of an EF-Tu recognizing the D-arm of a tRNA. The binding activity of EF-Tu1 was impaired by deletion of only 14 residues from the C-terminus, indicating that the C-terminus of EF-Tu1 is required for its binding to T-armless tRNA. These results suggest that C. elegans EF-Tu1 recognizes the D-arm instead of the T-arm by a mechanism involving its C-terminal region. This study sheds light on the co-evolution of RNA and RNA-binding proteins in nematode mitochondria.

Original languageEnglish
Pages (from-to)249-256
Number of pages8
JournalBiochemical Journal
Volume399
Issue number2
DOIs
Publication statusPublished - Oct 15 2006

Fingerprint

Peptide Elongation Factor Tu
Peptide Elongation Factors
Mitochondria
Transfer RNA
RNA
Proteins
Caenorhabditis elegans
RNA-Binding Proteins
Assays
Amino Acids

Keywords

  • Caenorhabditis elegans
  • Elongation factor-Tu (EF-Tu1)
  • Mitochondrion
  • Nematode
  • Translation
  • tRNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

A protein extension to shorten RNA : Elongated elongation factor-Tu recognizes the D-arm of T-armless tRNAs in nematode mitochondria. / Sakurai, Masayuki; Watanabe, Yoh Ichi; Watanabe, Kimitsuna; Ohtsuki, Takashi.

In: Biochemical Journal, Vol. 399, No. 2, 15.10.2006, p. 249-256.

Research output: Contribution to journalArticle

@article{4956f7b74c0c4136aac4a7bafbcbc0ee,
title = "A protein extension to shorten RNA: Elongated elongation factor-Tu recognizes the D-arm of T-armless tRNAs in nematode mitochondria",
abstract = "Nematode mitochondria possess extremely truncated tRNAs. Of 22 tRNAs, 20 lack the entire T-arm. The T-arm is necessary for the binding of canonical tRNAs and EF (elongation factor)-Tu (thermo-unstable). The nematode mitochondrial translation system employs two different EF-Tu factors named EF-Tu1 and EF-Tu2. Our previous study showed that nematode Caenorhabditis elegans EF-Tu1 binds specifically to T-armless tRNA. C. elegans EF-Tu1 has a 57-amino acid C-terminal extension that is absent from canonical EF-Tu, and the T-arm-binding residues of canonical EF-Tu are not conserved. In this study, the recognition mechanism of T-armless tRNA by EF-Tu1 was investigated. Both modification interference assays and primer extension analysis of cross-linked ternary complexes revealed that EF-Tu1 interacts not only with the tRNA acceptor stem but also with the D-arm. This is the first example of an EF-Tu recognizing the D-arm of a tRNA. The binding activity of EF-Tu1 was impaired by deletion of only 14 residues from the C-terminus, indicating that the C-terminus of EF-Tu1 is required for its binding to T-armless tRNA. These results suggest that C. elegans EF-Tu1 recognizes the D-arm instead of the T-arm by a mechanism involving its C-terminal region. This study sheds light on the co-evolution of RNA and RNA-binding proteins in nematode mitochondria.",
keywords = "Caenorhabditis elegans, Elongation factor-Tu (EF-Tu1), Mitochondrion, Nematode, Translation, tRNA",
author = "Masayuki Sakurai and Watanabe, {Yoh Ichi} and Kimitsuna Watanabe and Takashi Ohtsuki",
year = "2006",
month = "10",
day = "15",
doi = "10.1042/BJ20060781",
language = "English",
volume = "399",
pages = "249--256",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - A protein extension to shorten RNA

T2 - Elongated elongation factor-Tu recognizes the D-arm of T-armless tRNAs in nematode mitochondria

AU - Sakurai, Masayuki

AU - Watanabe, Yoh Ichi

AU - Watanabe, Kimitsuna

AU - Ohtsuki, Takashi

PY - 2006/10/15

Y1 - 2006/10/15

N2 - Nematode mitochondria possess extremely truncated tRNAs. Of 22 tRNAs, 20 lack the entire T-arm. The T-arm is necessary for the binding of canonical tRNAs and EF (elongation factor)-Tu (thermo-unstable). The nematode mitochondrial translation system employs two different EF-Tu factors named EF-Tu1 and EF-Tu2. Our previous study showed that nematode Caenorhabditis elegans EF-Tu1 binds specifically to T-armless tRNA. C. elegans EF-Tu1 has a 57-amino acid C-terminal extension that is absent from canonical EF-Tu, and the T-arm-binding residues of canonical EF-Tu are not conserved. In this study, the recognition mechanism of T-armless tRNA by EF-Tu1 was investigated. Both modification interference assays and primer extension analysis of cross-linked ternary complexes revealed that EF-Tu1 interacts not only with the tRNA acceptor stem but also with the D-arm. This is the first example of an EF-Tu recognizing the D-arm of a tRNA. The binding activity of EF-Tu1 was impaired by deletion of only 14 residues from the C-terminus, indicating that the C-terminus of EF-Tu1 is required for its binding to T-armless tRNA. These results suggest that C. elegans EF-Tu1 recognizes the D-arm instead of the T-arm by a mechanism involving its C-terminal region. This study sheds light on the co-evolution of RNA and RNA-binding proteins in nematode mitochondria.

AB - Nematode mitochondria possess extremely truncated tRNAs. Of 22 tRNAs, 20 lack the entire T-arm. The T-arm is necessary for the binding of canonical tRNAs and EF (elongation factor)-Tu (thermo-unstable). The nematode mitochondrial translation system employs two different EF-Tu factors named EF-Tu1 and EF-Tu2. Our previous study showed that nematode Caenorhabditis elegans EF-Tu1 binds specifically to T-armless tRNA. C. elegans EF-Tu1 has a 57-amino acid C-terminal extension that is absent from canonical EF-Tu, and the T-arm-binding residues of canonical EF-Tu are not conserved. In this study, the recognition mechanism of T-armless tRNA by EF-Tu1 was investigated. Both modification interference assays and primer extension analysis of cross-linked ternary complexes revealed that EF-Tu1 interacts not only with the tRNA acceptor stem but also with the D-arm. This is the first example of an EF-Tu recognizing the D-arm of a tRNA. The binding activity of EF-Tu1 was impaired by deletion of only 14 residues from the C-terminus, indicating that the C-terminus of EF-Tu1 is required for its binding to T-armless tRNA. These results suggest that C. elegans EF-Tu1 recognizes the D-arm instead of the T-arm by a mechanism involving its C-terminal region. This study sheds light on the co-evolution of RNA and RNA-binding proteins in nematode mitochondria.

KW - Caenorhabditis elegans

KW - Elongation factor-Tu (EF-Tu1)

KW - Mitochondrion

KW - Nematode

KW - Translation

KW - tRNA

UR - http://www.scopus.com/inward/record.url?scp=33749983710&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749983710&partnerID=8YFLogxK

U2 - 10.1042/BJ20060781

DO - 10.1042/BJ20060781

M3 - Article

C2 - 16859488

AN - SCOPUS:33749983710

VL - 399

SP - 249

EP - 256

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -