Kozmin Stanislav G, Dominska Margaret, Kokoska Robert J, Petes Thomas D
Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Duke University, Durham, NC 27710, USA.
Biological and Biotechnology Sciences, DEVCOM-ARL Army Research Office, 800 Park Offices Drive, Durham, NC 27703, USA.
Genetics. 2025 Jan 8;229(1):1-31. doi: 10.1093/genetics/iyae194.
Near the C-terminus of histone H2A in the yeast Saccharomyces cerevisiae, there are 2 serines (S122 and S129) that are targets of phosphorylation. The phosphorylation of serine 129 in response to DNA damage is dependent on the Tel1 and Mec1 kinases. In Schizosaccharomyces pombe and S. cerevisiae, the phosphorylation of serine 122 is dependent on the Bub1 kinase, and S. pombe strains with an alanine mutation of this serine have elevated levels of lagging chromosomes in mitosis. Strains that lack both Tel1 and Mec1 in S. cerevisiae have very elevated rates of nondisjunction. To clarify the functional importance of phosphorylation of serines 122 and 129 in H2A, we measured chromosome loss rates in single-mutant strains and double-mutant combinations. We also examined the interaction of mutations of BUB1, TEL1, and MEC1 in combination with mutations of serines 122 and 129 in H2A. We conclude that the phosphorylation state of S129 has no effect on chromosome disjunction whereas mutations that inactivate Bub1 or a S122A mutation in the histone H2A greatly elevate the rate of chromosome nondisjunction. Based on this analysis, we suggest that Bub1 exerts its primary effect on chromosome disjunction by phosphorylating S122 of histone H2A. However, Tel1, Mec1, and Bub1 are also functionally redundant in a second pathway affecting chromosome disjunction that is at least partially independent of phosphorylation of S122 of H2A.
在酿酒酵母中,组蛋白H2A的C末端附近有两个丝氨酸(S122和S129)是磷酸化的靶点。响应DNA损伤时丝氨酸129的磷酸化依赖于Tel1和Mec1激酶。在粟酒裂殖酵母和酿酒酵母中,丝氨酸122的磷酸化依赖于Bub1激酶,该丝氨酸发生丙氨酸突变的粟酒裂殖酵母菌株在有丝分裂中滞后染色体水平升高。酿酒酵母中同时缺失Tel1和Mec1的菌株不分离率非常高。为了阐明H2A中丝氨酸122和129磷酸化的功能重要性,我们测量了单突变菌株和双突变组合中的染色体丢失率。我们还研究了BUB1、TEL1和MEC1的突变与H2A中丝氨酸122和129的突变的相互作用。我们得出结论,S129的磷酸化状态对染色体分离没有影响,而使Bub1失活的突变或组蛋白H2A中的S122A突变会大大提高染色体不分离率。基于此分析,我们认为Bub1通过磷酸化组蛋白H2A的S122对染色体分离发挥主要作用。然而,Tel1、Mec1和Bub1在影响染色体分离的第二条途径中在功能上也是冗余的,该途径至少部分独立于H2A的S122的磷酸化。