Plant pre-mRNA splicing in fission yeast, Schizosaccharomyces pombe

Sarmah, Bhaskarjyoti; Chakraborty, Niranjan; Chakraborty, Subhra; Datta, Asis

doi:10.1016/s0006-291x(02)00364-9

Cited by 8 publications

(6 citation statements)

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“…The protein was synthesized during the grain development like other storage proteins . It was one of the first characterized protein encoded by a nonallergenic seed storage albumin gene. ,,, In addition, we and others have shown that AmA1 regulates the biology of nutrition, and its ectopic expression in storage organs such as tuber and seed helps increase protein and amino acid accumulation. , Further, AmA1 plays an important physiological role during organ development and is crucial for the homoblastic growth of potato tuber, the gradual transition cascade from stolon to mature tuber. , Thus we earlier hypothesized that induction of protein synthesis or mitogenic activity might be the result of overexpression of AmA1 or its turned-over products as signal molecule . In a recent study, our preliminary data suggested that proteome rebalancing due to AmA1 expression might lead to nutrient enhancement and increased yield .…”

Section: Introductionmentioning

confidence: 99%

Comparative Proteomics Reveals a Role for Seed Storage Protein AmA1 in Cellular Growth, Development, and Nutrient Accumulation

et al. 2013

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Seed storage proteins are known to be utilized as carbon and nitrogen source for growing seedlings and thus are considered as potential candidates for nutritional improvement. However, their precise function remains unknown. We have earlier shown that ectopic expression of a seed storage protein, AmA1, leads to increase in protein besides high tuber yield in potato. To elucidate the AmA1-regulated molecular mechanism affecting increased protein synthesis, reserve accumulation, and enhanced growth, a comparative proteomics approach has been applied to tuber life-cycle between wild-type and AmA1 potato. The differential display of proteomes revealed 150 AmA1-responsive protein spots (ARPs) that change their intensities more than 2.5-fold. The LC-ESI-MS/MS analyses led to the identification of 80 ARPs presumably associated with cell differentiation, regulating diverse functions, viz., protein biogenesis and storage, bioenergy and metabolism, and cell signaling. Metabolome study indicated up-regulation of amino acids paralleling the proteomics analysis. To validate this, we focused our attention on anatomical study that showed differences in cell size in the cortex, premedullary zone and pith of the tuber, coinciding with AmA1 expression and localization. Further, we interrogated the proteome data using one-way analysis of variance, cluster, and partial correlation analysis that identified two significant protein modules and six small correlation groups centered around isoforms of cysteine protease inhibitor, actin, heat shock cognate protein 83 and 14-3-3, pointing toward AmA1-regulated overlapping processes of protein enhancement and cell growth perhaps through a common mechanism of function. A model network was constructed using the protein data sets, which aim to show how target proteins might work in coordinated fashion and attribute to increased protein synthesis and storage reserve accumulation in AmA1 tubers on one hand and organ development on the other.

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Section: Introductionmentioning

confidence: 99%

Comparative Proteomics Reveals a Role for Seed Storage Protein AmA1 in Cellular Growth, Development, and Nutrient Accumulation

et al. 2013

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“…However, only a few plant genes were reported to be expressed in S. pombe (5)(6)(7)(8)(9). In addition, Sarmah et al, (10) also showed the successful splicing of a plant pre-mRNA by S. pombe (10).…”

Section: Introductionmentioning

confidence: 99%

Expression of Plant β-Glucosidase Genes inSchizosaccharomyces Pombe

Palabıyık

Esen

Albayrak

et al. 2006

Biotechnology & Biotechnological Equipment

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Schizosaccharomyces pombe as a model host organism was used for the functional analysis of plant genes. The heterologous expressions of dhr1 and myr1 genes encoding β-glucosidase in Sorghum bicolor and Sinapis alba, respectively, have been investigated in this fission yeast. cDNA's of these genes are cloned in pART1 and pREP42 expression vectors and transferred to S. pombe leu1-32 and ura4-D18 strains by electroporation. The transformants are screened by using colony PCR to identify the colonies containing plant genes. It was determined that pART1 was not a suitable vector for cloning these genes. Therefore, further analyses were done using recombinant vectors derived from pREP42. Transcripts of the genes from the recombinant clones were detected by RT-PCR using gene specific primers. Crude protein extracts prepared from these clones were analyzed by SDS-PAGE and Western blotting. In addition, these extracts were assayed for the enzyme activity. It was determined that plant β-glucosidase genes were expressed at transcriptional and translational levels in S. pombe, but their protein products were inactive. These results may indicate some deficiencies in the post translational modifications or negative interactions between S. pombe and plant β-glucosidases.

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“…Importantly, a single bona fide member of the serine/arginine-rich (SR) family of splicing regulators and several SR-like proteins implicated in the regulation of alternative splicing are encoded within the S. pombe genome (22,23). Furthermore, cis-acting sequence elements important in splicing regulation in multicellular eukaryotes have been shown to modulate the splicing efficiency of individual S. pombe introns (24), and indeed nonendogenous plant and mammalian intron sequences have been shown to be properly excised in S. pombe (25,26). Nevertheless, although instances of intron-retention have been documented (27,28), no examples of exon skipping have been described in S. pombe.…”

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Lariat sequencing in a unicellular yeast identifies regulated alternative splicing of exons that are evolutionarily conserved with humans

Awan

Manfredo

Pleiss

2013

Proc. Natl. Acad. Sci. U.S.A.

114

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Alternative splicing is a potent regulator of gene expression that vastly increases proteomic diversity in multicellular eukaryotes and is associated with organismal complexity. Although alternative splicing is widespread in vertebrates, little is known about the evolutionary origins of this process, in part because of the absence of phylogenetically conserved events that cross major eukaryotic clades. Here we describe a lariat-sequencing approach, which offers high sensitivity for detecting splicing events, and its application to the unicellular fungus, Schizosaccharomyces pombe, an organism that shares many of the hallmarks of alternative splicing in mammalian systems but for which no previous examples of exon-skipping had been demonstrated. Over 200 previously unannotated splicing events were identified, including examples of regulated alternative splicing. Remarkably, an evolutionary analysis of four of the exons identified here as subject to skipping in S. pombe reveals high sequence conservation and perfect length conservation with their homologs in scores of plants, animals, and fungi. Moreover, alternative splicing of two of these exons have been documented in multiple vertebrate organisms, making these the first demonstrations of identical alternative-splicing patterns in species that are separated by over 1 billion y of evolution.pre-mRNA splicing | post-transcriptional gene regulation | phylogeny T he protein coding regions of eukaryotic genes are typically interrupted by noncoding introns that must be removed to produce a translatable mRNA. The removal of introns, catalyzed by the spliceosome, offers a powerful opportunity for an organism to regulate gene expression. In mammals, where individual genes are often interrupted by multiple introns, it is now abundantly clear that the process of intron removal provides a critical regulatory control point for both qualitative and quantitative aspects of gene expression (1). By changing the identity of the exons that are included within the final mRNA, the process of alternative splicing plays a critical role in expanding the diversity of proteins that can be synthesized within a cell (2). Moreover, alternative splicing can direct the production of isoforms of genes that are directly targeted to cellular decay pathways, providing a mechanism to quantitatively regulate gene expression (3, 4).In mammalian organisms the predominant form of alternative splicing is exon skipping, wherein different combinations of exons are included in the final transcript. In contrast, exon skipping is far less prevalent in simpler eukaryotes (5, 6); however, recent studies suggest that splicing in the last eukaryotic common ancestor was similar in many respects to splicing in vertebrates, in so much as it was intron-dense (7-9), had degenerate splice site sequences (10), and likely had many of the proteins involved in alternative splicing (11,12). Intron density correlates positively with the prevalence of alternative splicing across the eukaryotic kingdoms (13), and thus it has...

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Plant pre-mRNA splicing in fission yeast, Schizosaccharomyces pombe

Cited by 8 publications

References 26 publications

Comparative Proteomics Reveals a Role for Seed Storage Protein AmA1 in Cellular Growth, Development, and Nutrient Accumulation

Comparative Proteomics Reveals a Role for Seed Storage Protein AmA1 in Cellular Growth, Development, and Nutrient Accumulation

Expression of Plant β-Glucosidase Genes inSchizosaccharomyces Pombe

Lariat sequencing in a unicellular yeast identifies regulated alternative splicing of exons that are evolutionarily conserved with humans

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