Ribosome binding sites on chloroplast rbcL and psbA mRNAs and light induced initiation of D1 translation英文文献.pdfVIP

Plant Molecular Biology 25: 437-448, 1994. © 1994 Kluwer Academic Publishers. Printed in Belgium. 437 Ribosome-binding sites on chloroplast rbcL and psbA mRNAs and light-induced initiation of D1 translation Jungmook Kim and John E. Mullet* Department of Biochemistry and Biophysics, Texas A M University, College Station, TX 77843, USA (* authorfor correspondence) Received 14 December 1993; accepted in revised form 4 March 1994 Key words: chloroplast, light induction, psbA, rbcL, translation initiation Abstract Chloroplast ribosome-binding sites were identified on the plastid rbcL and psbA mRNAs using toeprint analysis. The rbcL translation initiation domain is highly conserved and contains a prokaryotic Shine- Dalgarno (SD) sequence (GGA G G ) located 4 to 12 nucleotides upstream of the initiator AUG. Toeprint analysis ofrbcL m R N A associated with plastid polysomes revealed strong toeprint signals 15 nucleotides downstream from the A U G indicating ribosome binding at the translation initiation site. Escherichia coli 30S ribosomes generated similar toeprint signals when mixed with rbcL m R N A in the presence of ini- tiator tRNA. These results indicate that plastid SD sequences are functional in chloroplast translation initiation. The psbA initiator region lacks a SD sequence within 12 nucleotides of the initiator AUG. However, toeprint analysis of soluble and membrane polysome-associated psbA m R N A revealed ribo- somes bound to the initiator region. E. coli 30S ribosomes did not associate with the psbA t