Functional role of the C-terminal tail of the archaeal ribosomal stalk in recruitment of two elongation factors to the sarcin/ricin loop of 23S rRNA

Hirotatsu Imai, Tomohiro Miyoshi, Ryo Murakami, Kosuke Ito, Yoshizumi Ishino, Toshio Uchiumi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Two types of elongation factors alternate in their binding to the factor-binding center of the ribosome. Both binding events are accompanied by GTP hydrolysis and drive the translation elongation cycle. The multicopy ribosomal protein family, termed the stalk, contributes actively to the elongation process. Recent evidence indicates that the mobile C-terminal tail of archaeal stalk aP1 directly interacts with both the elongation factors aEF1A and aEF2. To investigate the functional significance of these interactions in recruitment of elongation factors to the factor-binding center of the ribosome, we substituted the archaeal stalk complex aL10•aP1 for the bL10•bL12 stalk complex in the Escherichia coli 50S subunit. The resultant hybrid ribosome accessed archaeal aEF1A and aEF2 in a manner dependent on the C-terminal tail containing the hydrophobic residues Leu103, Leu106 and Phe107. Bases G2659 and A2660 in the sarcin/ricin loop (SRL) of 23S rRNA were protected against DMS modification by both factors as was A1067 by aEF2. Mutagenesis indicated that this protection was dependent on the intact C-terminal tail of aP1. The results suggest a crucial role for the interactions between the stalk C-terminal tail and elongation factors in their recruitment to the SRL of 23S rRNA within the ribosome.

Original languageEnglish
Pages (from-to)613-624
Number of pages12
JournalGenes to Cells
Volume20
Issue number7
DOIs
Publication statusPublished - Jul 1 2015

Fingerprint

Peptide Elongation Factors
Ricin
Ribosomes
Ribosomal Proteins
Guanosine Triphosphate
Mutagenesis
Hydrolysis
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

Cite this

Functional role of the C-terminal tail of the archaeal ribosomal stalk in recruitment of two elongation factors to the sarcin/ricin loop of 23S rRNA. / Imai, Hirotatsu; Miyoshi, Tomohiro; Murakami, Ryo; Ito, Kosuke; Ishino, Yoshizumi; Uchiumi, Toshio.

In: Genes to Cells, Vol. 20, No. 7, 01.07.2015, p. 613-624.

Research output: Contribution to journalArticle

Imai, Hirotatsu ; Miyoshi, Tomohiro ; Murakami, Ryo ; Ito, Kosuke ; Ishino, Yoshizumi ; Uchiumi, Toshio. / Functional role of the C-terminal tail of the archaeal ribosomal stalk in recruitment of two elongation factors to the sarcin/ricin loop of 23S rRNA. In: Genes to Cells. 2015 ; Vol. 20, No. 7. pp. 613-624.
@article{766189b2443b454c8c838351753c6761,
title = "Functional role of the C-terminal tail of the archaeal ribosomal stalk in recruitment of two elongation factors to the sarcin/ricin loop of 23S rRNA",
abstract = "Two types of elongation factors alternate in their binding to the factor-binding center of the ribosome. Both binding events are accompanied by GTP hydrolysis and drive the translation elongation cycle. The multicopy ribosomal protein family, termed the stalk, contributes actively to the elongation process. Recent evidence indicates that the mobile C-terminal tail of archaeal stalk aP1 directly interacts with both the elongation factors aEF1A and aEF2. To investigate the functional significance of these interactions in recruitment of elongation factors to the factor-binding center of the ribosome, we substituted the archaeal stalk complex aL10•aP1 for the bL10•bL12 stalk complex in the Escherichia coli 50S subunit. The resultant hybrid ribosome accessed archaeal aEF1A and aEF2 in a manner dependent on the C-terminal tail containing the hydrophobic residues Leu103, Leu106 and Phe107. Bases G2659 and A2660 in the sarcin/ricin loop (SRL) of 23S rRNA were protected against DMS modification by both factors as was A1067 by aEF2. Mutagenesis indicated that this protection was dependent on the intact C-terminal tail of aP1. The results suggest a crucial role for the interactions between the stalk C-terminal tail and elongation factors in their recruitment to the SRL of 23S rRNA within the ribosome.",
author = "Hirotatsu Imai and Tomohiro Miyoshi and Ryo Murakami and Kosuke Ito and Yoshizumi Ishino and Toshio Uchiumi",
year = "2015",
month = "7",
day = "1",
doi = "10.1111/gtc.12256",
language = "English",
volume = "20",
pages = "613--624",
journal = "Genes to Cells",
issn = "1356-9597",
publisher = "Wiley-Blackwell",
number = "7",

}

TY - JOUR

T1 - Functional role of the C-terminal tail of the archaeal ribosomal stalk in recruitment of two elongation factors to the sarcin/ricin loop of 23S rRNA

AU - Imai, Hirotatsu

AU - Miyoshi, Tomohiro

AU - Murakami, Ryo

AU - Ito, Kosuke

AU - Ishino, Yoshizumi

AU - Uchiumi, Toshio

PY - 2015/7/1

Y1 - 2015/7/1

N2 - Two types of elongation factors alternate in their binding to the factor-binding center of the ribosome. Both binding events are accompanied by GTP hydrolysis and drive the translation elongation cycle. The multicopy ribosomal protein family, termed the stalk, contributes actively to the elongation process. Recent evidence indicates that the mobile C-terminal tail of archaeal stalk aP1 directly interacts with both the elongation factors aEF1A and aEF2. To investigate the functional significance of these interactions in recruitment of elongation factors to the factor-binding center of the ribosome, we substituted the archaeal stalk complex aL10•aP1 for the bL10•bL12 stalk complex in the Escherichia coli 50S subunit. The resultant hybrid ribosome accessed archaeal aEF1A and aEF2 in a manner dependent on the C-terminal tail containing the hydrophobic residues Leu103, Leu106 and Phe107. Bases G2659 and A2660 in the sarcin/ricin loop (SRL) of 23S rRNA were protected against DMS modification by both factors as was A1067 by aEF2. Mutagenesis indicated that this protection was dependent on the intact C-terminal tail of aP1. The results suggest a crucial role for the interactions between the stalk C-terminal tail and elongation factors in their recruitment to the SRL of 23S rRNA within the ribosome.

AB - Two types of elongation factors alternate in their binding to the factor-binding center of the ribosome. Both binding events are accompanied by GTP hydrolysis and drive the translation elongation cycle. The multicopy ribosomal protein family, termed the stalk, contributes actively to the elongation process. Recent evidence indicates that the mobile C-terminal tail of archaeal stalk aP1 directly interacts with both the elongation factors aEF1A and aEF2. To investigate the functional significance of these interactions in recruitment of elongation factors to the factor-binding center of the ribosome, we substituted the archaeal stalk complex aL10•aP1 for the bL10•bL12 stalk complex in the Escherichia coli 50S subunit. The resultant hybrid ribosome accessed archaeal aEF1A and aEF2 in a manner dependent on the C-terminal tail containing the hydrophobic residues Leu103, Leu106 and Phe107. Bases G2659 and A2660 in the sarcin/ricin loop (SRL) of 23S rRNA were protected against DMS modification by both factors as was A1067 by aEF2. Mutagenesis indicated that this protection was dependent on the intact C-terminal tail of aP1. The results suggest a crucial role for the interactions between the stalk C-terminal tail and elongation factors in their recruitment to the SRL of 23S rRNA within the ribosome.

UR - http://www.scopus.com/inward/record.url?scp=84934435240&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84934435240&partnerID=8YFLogxK

U2 - 10.1111/gtc.12256

DO - 10.1111/gtc.12256

M3 - Article

C2 - 26033302

AN - SCOPUS:84934435240

VL - 20

SP - 613

EP - 624

JO - Genes to Cells

JF - Genes to Cells

SN - 1356-9597

IS - 7

ER -