2‐D DIGE analysis of the budding yeast pH 6–11 proteome in meiosis

Abstract

Meiosis is the cell division that generates haploid gametes from diploid precursors. To provide insight into the functional proteome of budding yeast during meiosis, a 2-D DIGE kinetic approach was used to study proteins in the pH 6-11 range. Nearly 600 protein spots were visualised and 79 spots exhibited statistically significant changes in abundance as cells progressed through meiosis. Expression changes of up to 41-fold were detected and protein sequence information was obtained for 48 spots. Single protein identifications were obtained for 21 spots including different gel mobility forms of 5 proteins. A large number of post-translational events are suggested for these proteins, including processing, modification and import. The data are incorporated into an online 2-DE map of meiotic proteins in budding yeast, which extends our initial DIGE investigation of proteins in the pH 4-7 range. Together, the analyses provide peptide sequence data for 84 protein spots, including 50 single-protein identifications and gel mobility isoforms of 8 proteins. The largest classes of identified proteins include carbon metabolism, protein catabolism, protein folding, protein synthesis and the oxidative stress response. A number of the corresponding genes are required for yeast meiosis and recent studies have identified similar classes of proteins expressed during mammalian meiosis. This proteomic investigation and the resulting protein reference map make an important contribution towards a more detailed molecular view of yeast meiosis.

Dma1-dependent degradation of Septation Initiation Network proteins during meiosis inSchizosaccharomyces pombe

“…Targeted stage-specific proteolysis of cyclins and other proteins is already established as important for meiotic progression in S. pombe (Blanco et al, 2001;Funaya et al, 2012;Izawa et al, 2005;Kimata et al, 2008;Okuzaki et al, 2010;Pérez-Hidalgo et al, 2008). Furthermore, studies in Saccharomyces cerevisiae using 2D gels have revealed some changes in protein level during meiosis Scaife et al, 2010). We have shown in this study that Dma1p activity is required for the degradation of proteins at different stages of meiosis.…”

Distinct temporal requirements for autophagy and the proteasome in yeast meiosis

“…That study found that 142 protein spots were temporally regulated during meiosis but only identified 44 unique proteins by LC-MS/MS. 58,59 Here, TMT-based quantitative proteomics was applied to highly synchronized yeast cells at successive intervals following transfer to SPM, and 381 differentially expressed proteins (1.5-fold) of 2376 total identified proteins were found, significantly expanding our knowledge of meiosis at the proteome level. According to the studies of Chu et al and Primig et al, more than 1000 genes among approximately 6200 protein-encoding yeast genes showed significant changes (either induction or repression) in mRNA levels during sporulation.…”

Proteomic identification of germline proteins inCaenorhabditis elegans

“…Similar results have been seen in other studies-of the 50 proteins identified as being part of the yeast meiosis proteome, 30% are mitochondrial in function. 18,19 This is not surprising: in adult worms, the majority of cell growth and division, which are both energy-intensive processes, takes place in the germline. [20][21][22] Thus, the abundance of mitochondrial proteins in adult worms will correlate with the presence of a germline.…”