Abstract
Key message
Processed chloroplast RNAs are co-enriched with preparations of the chloroplast transcriptionally active chromosome.
Abstract
Chloroplast genomes are organized as a polyploid DNA–protein structure called the nucleoid. Transcriptionally active chloroplast DNA together with tightly bound protein factors can be purified by gel filtration as a functional entity called the transcriptionally active chromosome (TAC). Previous proteomics analyses of nucleoids and of TACs demonstrated a considerable overlap in protein composition including RNA binding proteins. Therefore the RNA content of TAC preparations from Nicotiana tabacum was determined using whole genome tiling arrays. A large number of chloroplast RNAs was found to be associated with the TAC. The pattern of RNAs attached to the TAC consists of RNAs produced by different chloroplast RNA polymerases and differs from the pattern of RNA found in input controls. An analysis of RNA splicing and RNA editing of selected RNA species demonstrated that TAC-associated RNAs are processed to a similar extent as the RNA in input controls. Thus, TAC fractions contain a specific subset of the processed chloroplast transcriptome.
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Acknowledgements
Generous funding by the Deutsche Forschungsgemeinschaft (Grant No. SCHM 1698/5-1) and the Einstein foundation (Einstein Stiftung Berlin) to CSL is gratefully acknowledged. Support of M.-K. L. by the IRI for Life Sciences is gratefully acknowledged.
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CSL and KK conception and design of study, MKL, SF, JM, SO acquisition of data, MKL, SF, CSL and KK analysis and/or interpretation of data, CSL drafting the manuscript, KK, MKL, SF, JM revising the manuscript critically for important intellectual content and CSL, KK, MKL, SF, JM, SO approval of the version of the manuscript to be published.
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Lehniger, MK., Finster, S., Melonek, J. et al. Global RNA association with the transcriptionally active chromosome of chloroplasts. Plant Mol Biol 95, 303–311 (2017). https://doi.org/10.1007/s11103-017-0649-x
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DOI: https://doi.org/10.1007/s11103-017-0649-x