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有丝分裂的蛋白质组学分析及裂殖酵母中新的短开放阅读框的特征描述。

Proteomic analysis of meiosis and characterization of novel short open reading frames in the fission yeast .

机构信息

Department of Genetics, Faculty of Natural Sciences, Comenius University in Bratislava , Bratislava, Slovakia.

Department of Membrane Biochemistry, Inst. Of Animal Biochemistry and Genetics, Centre of Biosciences, Slovak Academy of Sciences , Bratislava, Slovakia.

出版信息

Cell Cycle. 2020 Jul;19(14):1777-1785. doi: 10.1080/15384101.2020.1779470. Epub 2020 Jun 17.

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 . 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

减数分裂是由二倍体前体细胞产生单倍体配子的过程。我们使用稳定同位素标记的细胞培养中的氨基酸(SILAC)来描述 裂殖酵母中的减数分裂蛋白质组。我们比较了在减数分裂 I 周围收获的细胞中提取的蛋白质的相对水平与减数分裂 II 中的蛋白质水平,以及减数分裂间隔期间没有 S 期的有丝分裂前期 S 期的蛋白质水平。我们的蛋白质组数据集揭示了与核糖体图谱先前鉴定的短开放阅读框(sORF)相对应的肽,这些 sORF 被鉴定为新的翻译区域。我们通过 Western blot 验证了选定 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一个资源,这可能有助于理解减数分裂 I 和减数分裂 II 之间的差异,以及如何在两个减数分裂之间抑制 S 期。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/faf9/7469465/441d5c20496d/KCCY_A_1779470_F0001_OC.jpg

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