Identification of an OPR protein involved in the translation initiation of the PsaB subunit of photosystem I
Article first published online: 13 SEP 2012
© 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd
The Plant Journal
Volume 72, Issue 4, pages 652–661, November 2012
How to Cite
Rahire, M., Laroche, F., Cerutti, L. and Rochaix, J.-D. (2012), Identification of an OPR protein involved in the translation initiation of the PsaB subunit of photosystem I. The Plant Journal, 72: 652–661. doi: 10.1111/j.1365-313X.2012.05111.x
- Issue published online: 2 NOV 2012
- Article first published online: 13 SEP 2012
- Accepted manuscript online: 20 JUL 2012 11:41AM EST
- Received 6 June 2012; revised 29 June 2012; accepted 16 July 2012; published online 13 September 2012.
Figure S1. Recombinant MBP-Tab1 protein (aminoacids 390–747). A shorter version of Tab3 (amino acids 390– 747) fused to the maltose binding protein (MBP) wasproduced in E.coli with the pST4-MBP-psaB plasmid. Total proteins before and after 1 h of induction with 1mM IPTG (isopropyl β-D-1-thiogalactopyranoside) werefractionated by PAGE and stained with Coomassie Blue. The solublecell extract was loaded on an amylose resin (Biolabs), washed with20 mM Tris-HCl, pH 7.4, 0.2 M NaCl, 10 mM β-mercaptoethanol, 1mM EDTA and eluted with washing buffer containing 10 mM maltose.The four fractions from the elution are labeled 1 – 4. Fraction 2 was used for the RNA binding experiments.
Figure S2. Mutation of tab1-F15. Thefirst base of the CTG codon of L670 is missing in tab1-F15.This deletion creates the stop codon TGA.
Figure S3. Estimation of the amount of Tab1protein. Left: Recombinant Tab1 protein (Rec-Tab1) wasfractionated in decreasing amounts (50, 20, 10, 5, 2.5, 1 and 0 ng)together with 200 µg of total protein fromtab1-F15 by PAGE. Right: Decreasing amounts of totalprotein form wild type (200, 175, 159, 100, 50 and 0 µg) werefractionated with proteins from tab1-F15 so that thetotal amount of protein per lane was 200 µg. After blottingthe filter was reacted with Tab1 antibodies. From the autoradiogramit can be estimates that 2.5 ng of Rec-Tab1 yield a signal ofsimilar strength as that obtained with 200 µg ofwild-type proteins. The amount of Tab1 was estimated asindicated in the text. Authentic Tab1 is indicated with an arrowand recombinant Tab1 protein with a bar. M, size markers.
Table S1. Oligonucleotides.
Table S2. Determination of the essential region of Tab1.
Appendix S1. Determination of the quantity of Tab1 protein.
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