Abstract

Abiotic stress, including drought, salinity, and temperature extremes, regulates gene expression at the transcriptional and posttranscriptional levels. Expression profiling of total messenger RNAs (mRNAs) from rice (Oryza sativa) leaves grown under stress conditions revealed that the transcript levels of photosynthetic genes are reduced more rapidly than others, a phenomenon referred to as stress-induced mRNA decay (SMD). By comparing RNA polymerase II engagement with the steady-state mRNA level, we show here that SMD is a posttranscriptional event. The SMD of photosynthetic genes was further verified by measuring the half-lives of the small subunit of Rubisco (RbcS1) and Chlorophyll a/b-Binding Protein1 (Cab1) mRNAs during stress conditions in the presence of the transcription inhibitor cordycepin. To discern any correlation between SMD and the process of translation, changes in total and polysome-associated mRNA levels after stress were measured. Total and polysome-associated mRNA levels of two photosynthetic (RbcS1 and Cab1) and two stress-inducible (Dehydration Stress-Inducible Protein1 and Salt-Induced Protein) genes were found to be markedly similar. This demonstrated the importance of polysome association for transcript stability under stress conditions. Microarray experiments performed on total and polysomal mRNAs indicate that approximately half of all mRNAs that undergo SMD remain polysome associated during stress treatments. To delineate the functional determinant(s) of mRNAs responsible for SMD, the RbcS1 and Cab1 transcripts were dissected into several components. The expressions of different combinations of the mRNA components were analyzed under stress conditions, revealing that both 3' and 5' untranslated regions are necessary for SMD. Our results, therefore, suggest that the posttranscriptional control of photosynthetic mRNA decay under stress conditions requires both 3' and 5' untranslated regions and correlates with differential polysome association.

Highlights

  • Abiotic stress, including drought, salinity, and temperature extremes, regulates gene expression at the transcriptional and posttranscriptional levels

  • MRNA levels of the light reaction genes Chlorophyll a/b-Binding Protein1 (Cab1), Plastocyanin (PCY), Cab26, OEE1, PSI-D and PSI-K and the dark reaction genes Rubisco Small Subunit1 (RbcS1), Rubisco Activase (RA), SEDP2ase, GAPDH, TK, and FBPase-P are rapidly reduced in response to both drought and salt stress conditions

  • Our results suggest that the stress-induced mRNA decay (SMD) of RbcS1 and Cab1 as well as the control of the stress-inducible Dehydration Stress-Inducible Protein1 (Dip1) and Salt-Induced Protein (SalT) genes are posttranscriptional events

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Summary

Introduction

Abiotic stress, including drought, salinity, and temperature extremes, regulates gene expression at the transcriptional and posttranscriptional levels. The SMD of photosynthetic genes was further verified by measuring the half-lives of the small subunit of Rubisco (RbcS1) and Chlorophyll a/b-Binding Protein (Cab1) mRNAs during stress conditions in the presence of the transcription inhibitor cordycepin. Total and polysome-associated mRNA levels of two photosynthetic (RbcS1 and Cab1) and two stress-inducible (Dehydration Stress-Inducible Protein and Salt-Induced Protein) genes were found to be markedly similar. This demonstrated the importance of polysome association for transcript stability under stress conditions. Suggest that the posttranscriptional control of photosynthetic mRNA decay under stress conditions requires both 39 and 59 untranslated regions and correlates with differential polysome association. Additional investigations of DST elements in Arabidopsis have revealed that unstable transcripts encoding proteins associated with circadian control possess DST elements in their 39 UTRs (Gutierrez et al, 2002; Lidder et al, 2005; Streatfield, 2007)

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