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The Arabidopsis splicing regulator SR45 confers salt tolerance in a splice isoform-dependent manner

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Functions of most splice isoforms that are generated by alternative splicing are unknown. We show that two splice variants that encode proteins differing in only eight amino acids have distinct functions in a stress response.

Abstract

Serine/arginine-rich (SR) and SR-like proteins, a conserved family of RNA binding proteins across eukaryotes, play important roles in pre-mRNA splicing and other post-transcriptional processes. Pre-mRNAs of SR and SR-like proteins undergo extensive alternative splicing in response to diverse stresses and produce multiple splice isoforms. However, the functions of most splice isoforms remain elusive. Alternative splicing of pre-mRNA of Arabidopsis SR45, which encodes an SR-like splicing regulator, generates two isoforms (long—SR45.1 and short—SR45.2). The proteins encoded by these two isoforms differ in eight amino acids. Here, we investigated the role of SR45 and its splice variants in salt stress tolerance. The loss of SR45 resulted in enhanced sensitivity to salt stress and changes in expression and splicing of genes involved in regulating salt stress response. Interestingly, only the long isoform (SR45.1) rescued the salt-sensitive phenotype as well as the altered gene expression and splicing patterns in the mutant. These results suggest that SR45 positively regulates salt tolerance. Furthermore, only the long isoform is required for SR45-mediated salt tolerance.

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Acknowledgements

This research was supported by a Grant from the National Science Foundation (ABI 0743097). We thank Dr. Xiao-Ning Zhang, St. Bonaventure University, St. Bonaventure, NY for providing the seeds of complemented lines; Dr. Salah and Dr. Palusa for their help on this Project; Dr. Prasad for his comments on the manuscript. MA was supported by a Ph.D. fellowship from the government of the Kingdom of Saudi Arabia.

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  1. Mohammed Albaqami

    Present address: Department of Biology, Faculty of Applied Science, Umm Al-Qura University, Mecca, 21955, Kingdom of Saudi Arabia

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  1. Department of Biology and Program in Cell and Molecular Biology, Colorado State University, Fort Collins, CO, 80523, USA

    Mohammed Albaqami, K. Laluk & Anireddy S. N. Reddy

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  1. Mohammed Albaqami
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  2. K. Laluk
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  3. Anireddy S. N. Reddy
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Contributions

ASNR conceived and supervised the study. MA, KL, and ASNR designed experiments and analyzed data. MA and KL performed experiments. MA and ASNR wrote the manuscript.

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Correspondence to Anireddy S. N. Reddy.

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Supplementary material 1 (JPEG 44 kb)

Figure S1: Ion content (mg/g dry weight) of WT,sr45and the complemented lines in plants grown in the presence and absence of NaCl. Three-day-old MS grown seedlings were transferred to MS or MS supplemented with 150 mM NaCl and allowed to grow for two weeks before harvesting. Harvested seedlings were rinsed once with an excess volume of 50 mM EDTA followed by 2 rinses with bi-distilled water. The oven-dried tissues were digested and used for ICP analysis. Data represent the mean ± SE from the average of three independent experiments. Significance was calculated using analysis of variance and Tukey’s test, the significance is indicated by letters associated with bars

Supplementary material 2 (PDF 83 kb)

Figure S2: Sequence alignment ofSOS4isoforms. The amplified products were resolved in a gel and different bands were purified and sequenced. Sequences of cDNAs (A) and predicted proteins (B) were aligned. (C) A schematic diagram showing alternatively spliced transcripts of SOS4. Alternative splicing of the first intron generated three splice isoforms. Primers used in PCR are presented in Suppl. Table 1

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Albaqami, M., Laluk, K. & Reddy, A.S.N. The Arabidopsis splicing regulator SR45 confers salt tolerance in a splice isoform-dependent manner. Plant Mol Biol 100, 379–390 (2019). https://doi.org/10.1007/s11103-019-00864-4

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