Global analyses of sumoylated proteins in Saccharomyces cerevisiae - Induction of protein sumoylation by cellular stresses

Ludwig Institute for Cancer Research, University of California-San Diego, La Jolla, 92093, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 08/2004; 279(31):32262-8. DOI: 10.1074/jbc.M404173200
Source: PubMed

ABSTRACT We have undertaken a global analysis of sumoylated proteins in Saccharomyces cerevisiae by tandem mass spectrometry. Exposure of cells to oxidative and ethanol stresses caused large increases in protein sumoylation. A large number of new sumoylated proteins were identified in untreated, hydrogen peroxide-treated, and ethanol-treated cells. These proteins are known to be involved in diverse cellular processes, including gene transcription, protein translation, DNA replication, chromosome segregation, metabolic processes, and stress responses. Additionally, the known enzymes, including E1, E2, and E3 of the sumoylation cascade were found to be auto-sumoylated. Taken together, these results show that protein sumoylation is broadly involved in many cellular functions and this mass spectrometry-based proteomic approach is useful in studying the regulation of protein sumoylation in the cells.

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Available from: Weidong Zhou, May 28, 2015
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    • "Here, the relevant Sumo target still needs to be identified. A good candidate is RNAPII, which was previously shown to be sumoylated, although the exact physiological significance remains unclear (Wohlschlegel et al. 2004; Zhou et al. 2004; Chen et al. 2009). Histones may also be Sumo substrates (Nathan et al. 2006). "
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    ABSTRACT: Transcription factors are abundant Sumo targets, yet the global distribution of Sumo along the chromatin and its physiological relevance in transcription are poorly understood. Using Saccharomyces cerevisiae, we determined the genome-wide localization of Sumo along the chromatin. We discovered that Sumo-enriched genes are almost exclusively involved in translation, such as tRNA genes and ribosomal protein genes (RPGs). Whole-genome expression analysis showed that Sumo positively regulates their transcription. We also discovered that the Sumo consensus motif at RPG promoters is identical to the DNA binding motif of the transcription factor Rap1. We demonstrate that Rap1 is a molecular target of Sumo and that sumoylation of Rap1 is important for cell viability. Furthermore, Rap1 sumoylation promotes recruitment of the basal transcription machinery, and sumoylation of Rap1 cooperates with the target of rapamycin kinase complex 1 (TORC1) pathway to promote RPG transcription. Strikingly, our data reveal that sumoylation of Rap1 functions in a homeostatic feedback loop that sustains RPG transcription during translational stress. Taken together, Sumo regulates the cellular translational capacity by promoting transcription of tRNA genes and RPGs. Published by Cold Spring Harbor Laboratory Press.
    Genome Research 03/2015; 25(6). DOI:10.1101/gr.185793.114 · 13.85 Impact Factor
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    • "Roughly about 500 substrate proteins in budding yeast have been identified as SUMO substrate proteins (Srikumar et al. 2013; Zhou et al. 2004). SUMO is essential for the viability of yeast and regulates diverse biological processes, including DNA repair, transcriptional regulation, protein localization, formation of subnuclear structures, and cell cycle progression (Praefcke et al. 2012; Psakhye and Jentsch 2012; Srikumar et al. 2013; Zhou et al. 2004). In addition, the discovery of SUMO-dependent ubiquitin ligases, such as Slx5-Slx8 and Ris1, has revealed that SUMO enhances protein degradation by binding with ubiquitin (Praefcke et al. 2012; Uzunova et al. 2007). "
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    ABSTRACT: The complex inhibitory effects of inhibitors present in lignocellulose hydrolysate suppress the ethanol fermentation of Saccharomyces cerevisiae. Although the interactive inhibitory effects play important roles in the actual hydrolysate, few studies have investigated glycolaldehyde, the key inhibitor of hot-compressed water-treated lignocellulose hydrolysate. Given this challenge, we investigated the interactive effects of mixed fermentation inhibitors, including glycolaldehyde. First, we confirmed that glycolaldehyde was the most potent inhibitor in the hydrolysate and exerted interactive inhibitory effects in combination with major inhibitors. Next, through genome-wide analysis and megavariate data modeling, we identified SUMOylation as a novel potential mechanism to overcome the combinational inhibitory effects of fermentation inhibitors. Indeed, overall SUMOylation was increased and Pgk1, which produces an ATP molecule in glycolysis by substrate-level phosphorylation, was SUMOylated and degraded in response to glycolaldehyde. Augmenting the SUMO-dependent ubiquitin system in the ADH1-expressing strain significantly shortened the lag phase of growth, released cells from G2/M arrest, and improved energy status and glucose uptake in the inhibitor-containing medium. In summary, our study was the first to establish SUMOylation as a novel platform for regulating the lag phase caused by complex fermentation inhibitors.
    Applied Microbiology and Biotechnology 10/2014; 99(1). DOI:10.1007/s00253-014-6174-9 · 3.81 Impact Factor
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    • "covalently linked to lysine residues in a process similar to ubiquitination and, in doing so, affect the conformation, protein-protein interaction, or subcellular location of the modified protein (Hickey et al., 2012; Ulrich, 2009). Although SUMOylation of yeast Hsp82 (yHsp90) and human Hsp90a (hHsp90a) has been reported previously (Panse et al., 2004; Pountney et al., 2008; Zhou et al., 2004), identification of individual SUMOylated lysine residues and their impact on Hsp90 function remains unexplored. Here, we report that SUMOylation of an N domain lysine conserved in both yeast and human Hsp90 initiates recruitment of Aha1 to the chaperone complex in cells. "
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    ABSTRACT: The stability and activity of numerous signaling proteins in both normal and cancer cells depends on the dimeric molecular chaperone heat shock protein 90 (Hsp90). Hsp90's function is coupled to ATP binding and hydrolysis and requires a series of conformational changes that are regulated by cochaperones and numerous posttranslational modifications (PTMs). SUMOylation is one of the least-understood Hsp90 PTMs. Here, we show that asymmetric SUMOylation of a conserved lysine residue in the N domain of both yeast (K178) and human (K191) Hsp90 facilitates both recruitment of the adenosine triphosphatase (ATPase)-activating cochaperone Aha1 and, unexpectedly, the binding of Hsp90 inhibitors, suggesting that these drugs associate preferentially with Hsp90 proteins that are actively engaged in the chaperone cycle. Importantly, cellular transformation is accompanied by elevated steady-state N domain SUMOylation, and increased Hsp90 SUMOylation sensitizes yeast and mammalian cells to Hsp90 inhibitors, providing a mechanism to explain the sensitivity of cancer cells to these drugs.
    Molecular cell 01/2014; 53(2):317-29. DOI:10.1016/j.molcel.2013.12.007 · 14.46 Impact Factor
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