Department of Biology, Gettysburg College, Gettysburg, PA 17325, USA.
Data Analytics, Genencor Technology Center, IFF, Palo Alto, CA, 94306, USA.
Genetics. 2022 Sep 30;222(2). doi: 10.1093/genetics/iyac130.
Aspergillus nidulans snxA, an ortholog of Saccharomyces cerevisiae Hrb1/Gbp2 messenger RNA shuttle proteins, is-in contrast to budding yeast-involved in cell cycle regulation, in which snxA1 and snxA2 mutations as well as a snxA deletion specifically suppress the heat sensitivity of mutations in regulators of the CDK1 mitotic induction pathway. snxA mutations are strongly cold sensitive, and at permissive temperature snxA mRNA and protein expression are strongly repressed. Initial attempts to identify the causative snxA mutations revealed no defects in the SNXA protein. Here, we show that snxA1/A2 mutations resulted from an identical chromosome I-II reciprocal translocation with breakpoints in the snxA first intron and the fourth exon of a GYF-domain gene, gyfA. Surprisingly, a gyfA deletion and a reconstructed gyfA translocation allele suppressed the heat sensitivity of CDK1 pathway mutants in a snxA+ background, demonstrating that 2 unrelated genes, snxA and gyfA, act through the CDK1-CyclinB axis to restrain the G2-M transition, and for the first time identifying a role in G2-M regulation for a GYF-domain protein. To better understand snxA1/A2-reduced expression, we generated suppressors of snxA cold sensitivity in 2 genes: (1) loss of the abundant nucleolar protein Nsr1/nucleolin bypassed the requirement for snxA and (2) loss of the Set2 histone H3 lysine36 (H3K36) methyltransferase or a nonmethylatable histone H3K36L mutant rescued hypomorphic snxA mutants by restoring full transcriptional proficiency, indicating that methylation of H3K36 acts normally to repress snxA transcription. These observations are in line with known Set2 functions in preventing excessive and cryptic transcription of active genes.
青霉 nidulans snxA,酿酒酵母 Hrb1/Gbp2 信使 RNA 穿梭蛋白的直系同源物,与芽殖酵母不同,它参与细胞周期调控,snxA1 和 snxA2 突变以及 snxA 缺失特异性抑制 CDK1 有丝分裂诱导途径调节因子的热敏感性。snxA 突变体对冷非常敏感,在允许温度下,snxA mRNA 和蛋白表达受到强烈抑制。最初试图鉴定导致 snxA 突变的原因,发现 SNXA 蛋白没有缺陷。在这里,我们表明 snxA1/A2 突变是由于染色体 I-II 相互易位引起的,断点位于 snxA 的第一个内含子和 GYF 结构域基因 gyfA 的第四个外显子。令人惊讶的是,gyfA 缺失和重建的 gyfA 易位等位基因在 snxA+背景下抑制了 CDK1 途径突变体的热敏感性,表明 2 个不相关的基因 snxA 和 gyfA 通过 CDK1-CyclinB 轴发挥作用,抑制 G2-M 转变,并且首次确定了 GYF 结构域蛋白在 G2-M 调控中的作用。为了更好地理解 snxA1/A2 表达减少,我们在 2 个基因中产生了 snxA 冷敏感性的抑制子:(1)丰富的核仁蛋白 Nsr1/nucleolin 的缺失绕过了对 snxA 的需求,(2)缺失组蛋白 H3 赖氨酸 36(H3K36)甲基转移酶 Set2 或不可甲基化的 H3K36L 突变体通过恢复完全转录活性来拯救功能减弱的 snxA 突变体,表明 H3K36 的甲基化正常发挥作用以抑制 snxA 转录。这些观察结果与 Set2 已知的功能一致,即防止活性基因的过度和隐蔽转录。