Ogino Keiko, Hirota Kouji, Matsumoto Seiji, Takeda Tadayuki, Ohta Kunihiro, Arai Ken-ichi, Masai Hisao
Genome Dynamics Project and Department of Integrated Life Science, Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, Tokyo 113-8613, Japan.
Proc Natl Acad Sci U S A. 2006 May 23;103(21):8131-6. doi: 10.1073/pnas.0602498103. Epub 2006 May 12.
Cdc7 kinase, conserved through evolution, is known to be essential for mitotic DNA replication. The role of Cdc7 in meiotic recombination was suggested in Saccharomyces cerevisiae, but its precise role has not been addressed. Here, we report that Hsk1, the Cdc7-related kinase in Schizosaccharomyces pombe, plays a crucial role during meiosis. In a hsk1 temperature-sensitive strain (hsk1-89), meiosis is arrested with one nucleus state before meiosis I in most of the cells and meiotic recombination frequency is reduced by one order of magnitude, whereas premeiotic DNA replication is delayed but is apparently completed. Strikingly, formation of meiotic dsDNA breaks (DSBs) are largely impaired in the mutant, and Hsk1 kinase activity is essential for these processes. Deletion of all three checkpoint kinases, namely Cds1, Chk1, and Mek1, does not restore DSB formation, meiosis, or Cdc2 activation, which is suppressed in hsk1-89, suggesting that these aberrations are not caused by known checkpoint pathways but that Hsk1 may regulate DSB formation and meiosis. Whereas transcriptional induction of some rec genes and horsetail movement are normal, chromatin remodeling at ade6-M26, a recombination hotspot, which is prerequisite for subsequent DSB formation at this locus, is not observed in hsk1-89. These results indicate unique and essential roles of Hsk1 kinase in the initiation of meiotic recombination and meiosis.
细胞分裂周期7激酶(Cdc7激酶)在进化过程中高度保守,已知其对有丝分裂DNA复制至关重要。酿酒酵母中曾提示Cdc7在减数分裂重组中发挥作用,但其确切作用尚未得到明确阐述。在此,我们报告粟酒裂殖酵母中与Cdc7相关的激酶Hsk1在减数分裂过程中起关键作用。在hsk1温度敏感型菌株(hsk1-89)中,大多数细胞在减数分裂I之前的单核状态下减数分裂停滞,减数分裂重组频率降低一个数量级,而减数分裂前的DNA复制虽有延迟但明显完成。引人注目的是,该突变体中减数分裂双链DNA断裂(DSB)的形成在很大程度上受损,且Hsk1激酶活性对这些过程至关重要。缺失所有三种检查点激酶,即Cds1、Chk1和Mek1,不能恢复DSB形成、减数分裂或Cdc2激活,而在hsk1-89中这些过程受到抑制,这表明这些异常并非由已知的检查点途径引起,而是Hsk1可能调节DSB形成和减数分裂。虽然一些rec基因的转录诱导和马尾运动正常,但在hsk1-89中未观察到ade6-M26(一个重组热点)处的染色质重塑,而染色质重塑是该位点后续形成DSB的先决条件。这些结果表明Hsk1激酶在减数分裂重组起始和减数分裂过程中具有独特且至关重要的作用。