ABSTRACT

Meiosis is the process by which haploid gametes are produced from diploid precursor cells. We used stable isotope labeling by amino acids in cell culture (SILAC) to characterize the meiotic proteome in the fission yeast Schizosaccharomyces pombe. We compared relative levels of proteins extracted from cells harvested around meiosis I with those of meiosis II, and proteins from premeiotic S phase with the interval between meiotic divisions, when S phase is absent. Our proteome datasets revealed peptides corresponding to short open reading frames (sORFs) that have been previously identified by ribosome profiling as new translated regions. We verified expression of selected sORFs by Western blotting and analyzed the phenotype of deletion mutants. Our data provide a resource for studying meiosis that may help understand differences between meiosis I and meiosis II and how S phase is suppressed between the two meiotic divisions.

摘要

减数分裂是从二倍体前体细胞产生单倍体配子的过程。我们使用细胞培养物中氨基酸的稳定同位素标记 (SILAC) 来表征裂殖酵母裂殖酵母中的减数分裂蛋白质组。我们比较了从减数分裂 I 和减数分裂 II 周围收集的细胞中提取的蛋白质的相对水平，以及当 S 期不存在时，从减数分裂前 S 期收集的蛋白质与减数分裂之间的间隔的蛋白质的相对水平。我们的蛋白质组数据集揭示了与短开放阅读框（sORF）相对应的肽，这些肽先前已通过核糖体分析鉴定为新的翻译区域。我们通过蛋白质印迹验证了所选 sORF 的表达并分析了缺失突变体的表型。我们的数据为研究减数分裂提供了资源，可能有助于了解减数分裂 I 和减数分裂 II 之间的差异以及两个减数分裂之间 S 期如何受到抑制。