ppGpp inhibits peptide elongation cycle of chloroplast translation system in vitro

Yuhta Nomura, Taito Takabayashi, Hiroshi Kuroda, Yasushi Yukawa, Kwanchanok Sattasuk, Mitsuru Akita, Akira Nozawa, Yuzuru Tozawa

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

Chloroplasts possess common biosynthetic pathways for generating guanosine 3′,5′-(bis)pyrophosphate (ppGpp) from GDP and ATP by RelA-SpoT homolog enzymes. To date, several hypothetical targets of ppGpp in chloroplasts have been suggested, but they remain largely unverified. In this study, we have investigated effects of ppGpp on translation apparatus in chloroplasts by developing in vitro protein synthesis system based on an extract of chloroplasts isolated from pea (Pisum sativum). The chloroplast extracts showed stable protein synthesis activity in vitro, and the activity was sensitive to various types of antibiotics. We have demonstrated that ppGpp inhibits the activity of chloroplast translation in dose-effective manner, as does the toxic nonhydrolyzable GTP analog guanosine 5′-(β,γ-imido)triphosphate (GDPNP). We further examined polyuridylic acid-directed polyphenylalanine synthesis as a measure of peptide elongation activity in the pea chloroplast extract. Both ppGpp and GDPNP as well as antibiotics, fusidic acid and thiostrepton, inhibited the peptide elongation cycle of the translation system, but GDP in the similar range of the tested ppGpp concentration did not affect the activity. Our results thus show that ppGpp directly affect the translation system of chloroplasts, as they do that of bacteria. We suggest that the role of the ppGpp signaling system in translation in bacteria is conserved in the translation system of chloroplasts.

Original languageEnglish
Pages (from-to)185-196
Number of pages12
JournalPlant Molecular Biology
Volume78
Issue number1-2
DOIs
Publication statusPublished - Jan 1 2012
Externally publishedYes

Keywords

  • Antibiotics
  • Chloroplast
  • Elongation
  • In vitro translation
  • Pea
  • ppGpp

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

Fingerprint Dive into the research topics of 'ppGpp inhibits peptide elongation cycle of chloroplast translation system in vitro'. Together they form a unique fingerprint.

Cite this