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Ribonuclease II preserves chloroplast RNA homeostasis by increasing mRNA decay rates, and cooperates with polynucleotide phosphorylase in 3′ end maturation
Article first published online: 18 OCT 2012
© 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd
The Plant Journal
Volume 72, Issue 6, pages 960–971, December 2012
How to Cite
Germain, A., Kim, S. H., Gutierrez, R. and Stern, D. B. (2012), Ribonuclease II preserves chloroplast RNA homeostasis by increasing mRNA decay rates, and cooperates with polynucleotide phosphorylase in 3′ end maturation. The Plant Journal, 72: 960–971. doi: 10.1111/tpj.12006
Arabidopsis seed stocks: SALK_013306 (pnp1-1), CS85511 (P184L), CS87183 (pnp1-3), CS86422 (S202N ), CS93456 (A263V ), SALK_090294 (rnr1-3).
- Issue published online: 6 DEC 2012
- Article first published online: 18 OCT 2012
- Accepted manuscript online: 13 OCT 2012 08:23AM EST
- Received 19 June 2012; revised 20 August 2012; accepted 29 August 2012; published online 18 October 2012.
Figure S1. Effects on 5S rRNA accumulation of combining pnp and rnr mutations.
qRT-PCR was used to measure the amount of sequences containing mature 5S rRNA. Reverse transcription was performed using 3’ qRT-PCR gene-specific primers as described in Sharwood et al. (2011). The results were normalized to actin, and standard error is shown. The amount of 5S rRNA was set to 1 for A263, which acts as a wild-type control. The data are derived from two biological replicates and three technical replicates for each genotype.
Figure S2. RNA phenotypes of single and double mutants. RNA gel blots were probed with the segments shown in the lower right corner, with their extents indicated by underlining on the schematics.
Figure S3. Polysomal loading in WT and rnr1-3. Polysomes were separated in sucrose gradients as shown across the top, and RNA from fractions was analyzed by gel blot.
Figure S4. Analysis of psbC, rbcL and psbA mRNA in ActD-treated WT and rnr1-3 plants. The top part of the figure shows RNA accumulation by gel blot, with numbers at the left corresponding to transcripts whose accumulation is graphed below, as described in the legend to Figure 7.
Figure S5. Analysis of additional chloroplast transcripts following ActD treatment in WT and rnr1-3 plants.
Figure S6. Accumulation of RBCS mRNA in ActD-treated WT and rnr1-3 plants, as described in the legend to Figure 7.
Figure S7. Analysis of mitochondrial cox1, atp9 and rps4 mRNAs in ActD-treated WT and rnr1-3 plants.
Table S1. Sequences of oligonucleotides primers used in this study.
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