Analysis of the budding yeast pH 4–7 proteome in meiosis

Abstract

Meiosis, the developmental programme generating haploid gametes from diploid precursors, requires two cell divisions and many innovations. In budding yeast, a large number of genes are expressed exclusively during meiosis while others are repressed compared to vegetative growth. Microarray analysis has shown that gene expression during meiosis is highly regulated, and has been used to classify yeast genes according to meiotic temporal expression pattern. In this study, we have begun to investigate the kinetics of meiotic protein expression using a proteomics approach. 2-D DIGE was used to characterise the temporal protein expression patterns of the budding yeast pH 4-7 proteome in meiosis. More than 1400 meiotic protein spots were visualised and at least 63 spots were temporally regulated during meiosis in a statistically significant manner. Gel spots with significant expression changes were excised and 26 unique proteins were identified using LC-MS/MS. The identified proteins could be classified into functional categories and the genes encoding a number of these were previously shown to be involved in yeast sporulation and meiosis. This data set was used to assemble the first differential 2-D PAGE map of budding yeast meiosis, which can be accessed through a web server. This work represents one of the first quantitative proteomic analyses of meiosis in yeast and will provide a valuable resource for future investigations.

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.…”

Cell Differentiation and Spatial Organization in Yeast Colonies: Role of Cell-Wall Integrity Pathway

“…Although sporulation is a response to nutrient starvation, paradoxically, it is also a program that requires a sizable energy investment (Ray et al 2013). Energy is required for chromosomes to replicate, pair, recombine, and segregate in meiosis; for spore-wall formation [reviewed in Kupiec et al (1997)]; and for the induction of hundreds of gene products, some to very high levels (Chu et al 1998;Primig et al 2000;Grassl et al 2010). In laboratory sporulation cultures, respiration of acetate provides the energy for sporulation, but it is less clear how wild yeast communities obtain this energy.…”

A clinically based protein discovery strategy to identify potential biomarkers of response to anti‐TNF‐α treatment of psoriatic arthritis

“…Protein spots were excised from the gels and digested using a protocol similar to that of Grassl et al, 2010 but without the use of Ziptips [63]). Briefly, the gel spots were destained, reduced and alkylated, then dehydrated with ACN.…”