Ribosomes pause at specific sites during synthesis of membrane-bound chloroplast reaction center protein D1

Jungmook Kim, Patricia Gamble Klein, John E. Mullet

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

Photosynthetic reaction center protein D1 contains five membrane-spanning α-helices which form binding sites for pheophytin, chlorophyll, carotenoids, quinone, Fe2+, and probably Mn2+. D1 translation intermediates of 15 to 28 kD were detected when isolated chloroplasts were pulse-labeled with [35S]methionine. The D1 translation intermediates were associated with membrane polysemes and can be chased into full length D1. The sites of translation pausing were determined by mapping the distribution of ribosomes on D1 mRNA using toeprint analysis. Clusters of toeprint signals were generated by D1 mRNA associated with membranes but not by D1 mRNA in nonpolysomal fractions of the soluble phase or phenol-extracted mRNA. The distribution of ribosomes on D1 mRNA determined by toeprint analysis was consistent with D1 translation intermediates observed with pulse-labeling. Ribosome pausing may facilitate co-translational binding of cofactors such as chlorophyll to D1 and aid the integration of D1 into thylakoid membranes.

Original languageEnglish (US)
Pages (from-to)14931-14938
Number of pages8
JournalJournal of Biological Chemistry
Volume266
Issue number23
StatePublished - 1991
Externally publishedYes

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Photosystem II Protein Complex
Ribosomes
Membranes
Messenger RNA
Chlorophyll
Pheophytins
Photosynthetic Reaction Center Complex Proteins
Thylakoids
Chloroplasts
Carotenoids
Phenol
Methionine
Labeling
Binding Sites
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ribosomes pause at specific sites during synthesis of membrane-bound chloroplast reaction center protein D1. / Kim, Jungmook; Klein, Patricia Gamble; Mullet, John E.

In: Journal of Biological Chemistry, Vol. 266, No. 23, 1991, p. 14931-14938.

Research output: Contribution to journalArticle

Kim, Jungmook ; Klein, Patricia Gamble ; Mullet, John E. / Ribosomes pause at specific sites during synthesis of membrane-bound chloroplast reaction center protein D1. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 23. pp. 14931-14938.
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