Leem S H, Ogawa H
Department of Biology, Faculty of Science, Osaka University, Japan.
Nucleic Acids Res. 1992 Feb 11;20(3):449-57. doi: 10.1093/nar/20.3.449.
The MRE4 gene was cloned by complementation of the defects of meiotic recombination and haploidization in an mre4-1 mutant. Disruption of MRE4 resulted in reduced meiotic recombination and spore inviability. The mre4 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass the reductional division. Analysis of meiotic DNA extracted from the mre4 mutant cells revealed that double-strand breaks occurred at the two sites of the HIS4-LEU2 recombination hot spot, but at a frequency of about 10-20% of the wild type. Northern blot analysis indicated that the MRE4 gene produces four transcripts of 1.63, 3.2, 4.0 and 6.2 kb. All of these transcripts are absent from mitotic cells and are meiotically induced. The DNA sequence of the MRE4 open reading frame predicts a 497-amino acids protein with a molecular mass of 56.8 kDa. The Mre4 protein contains highly conserved amino acid sequences found specifically in serine-threonine protein kinases. These results suggest that protein phosphorylation is required directly or indirectly for meiotic recombination.
通过互补mre4 - 1突变体减数分裂重组和单倍体化缺陷克隆了MRE4基因。MRE4的破坏导致减数分裂重组减少和孢子无活力。mre4孢子致死性可被spo13抑制,spo13是一种使细胞绕过减数分裂的突变。对从mre4突变体细胞中提取的减数分裂DNA的分析表明,双链断裂发生在HIS4 - LEU2重组热点的两个位点,但频率约为野生型的10 - 20%。Northern印迹分析表明,MRE4基因产生1.63、3.2、4.0和6.2 kb的四种转录本。所有这些转录本在有丝分裂细胞中不存在,而是在减数分裂时被诱导产生。MRE4开放阅读框的DNA序列预测有一个497个氨基酸的蛋白质,分子量为56.8 kDa。Mre4蛋白含有在丝氨酸 - 苏氨酸蛋白激酶中特有的高度保守氨基酸序列。这些结果表明蛋白质磷酸化直接或间接参与减数分裂重组。