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Mutation of Arabidopsis SME1 improves oxidative stress resilience

ABSTRACT: Alternative splicing is an important posttranscriptional gene regulatory process, acting in diverse adaptive and basal plant processes. Splicing of precursor-messenger RNA (pre-mRNA) is catalyzed by a dynamic ribonucleoprotein complex, termed the spliceosome. In a suppressor screen, we identified a nonsense mutation in the Sm protein SME1 to alleviate photorespiratory H2O2-dependent cell death in catalase-deficient plants. Furthermore, sme1-2 mutants showed increased tolerance to the superoxide-generating herbicide methyl viologen. mRNA-seq and proteomic analyses indicated a constitutive stress response and pre-mRNA splicing alterations in sme1-2 mutants under control conditions. With a pulldown experiment, using SME1 as a bait, we provide experimental evidence for proteins associated with Arabidopsis thaliana spliceosome complexes, by identifying almost 50 proteins homologous to mammalian spliceosome-associated proteins and propose four novel proteins previously not-reported or hypothesized to be part of plant spliceosome complexes. Mutation of one of these interactors, the Sm core assembly protein ICLN also resulted in a decreased sensitivity to methyl viologen. Taken together, we show that SME1 mutation and impaired snRNP assembly result in enhanced resilience to oxidative stress.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

SUBMITTER: Patrick Willems

LAB HEAD: Frank Van Breusegem

PROVIDER: PXD024816 | Pride | 2023-03-03

REPOSITORIES: pride

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			Action	DRS
	H080428_45cm_CMB-329_KRGEV_PatrickW_KO-1.raw	Raw
	H080432_45cm_CMB-329_KRGEV_PatrickW_WT-1.raw	Raw
	H080434_45cm_CMB-329_KRGEV_PatrickW_KO-2.raw	Raw
	H080438_45cm_CMB-329_KRGEV_PatrickW_WT-2.raw	Raw
	H080440_45cm_CMB-329_KRGEV_PatrickW_KO-3.raw	Raw

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Arabidopsis SME interactome reveals spliceosome components

Project description:Alternative splicing (AS) can produce multiple transcripts with different exon-intron structures from a single pre-mRNA. Pre-mRNA splicing is catalyzed by a dynamic macromolecular ribonucleoprotein (RNP) complex termed the spliceosome. The spliceosome consists of several small nuclear ribonucleoproteins (snRNPs) that bind uridine-rich small nuclear RNA (snRNA). In U1, U2, U4 and U5 snRNPs, snRNA interacts with the conserved Smith antigen (Sm) proteins via a bipartite Sm sequence motif. In eukaryotes, seven Sm proteins (B, D1/2/3, E, F and G) form a heptameric ring-shaped complex surrounding the snRNA. Here, we performed a tandem affinity purification using SMEB as a bait in Arabidopsis cell suspension cultures. At least 45 known/hypothesized and potential novel spliceosome components were identified in Arabidopsis.

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