Fox M E, Yamada T, Ohta K, Smith G R
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Genetics. 2000 Sep;156(1):59-68. doi: 10.1093/genetics/156.1.59.
The heptamer sequence ATGACGT is essential for activity of the M26 meiotic recombination hotspot in the ade6 gene of Schizosaccharomyces pombe. Hotspot activity is associated with binding of the heterodimeric transcription factor Atf1.Pcr1 to M26. We have found that the sequences (C/T/G) TGACGT also bound Atf1.Pcr1 and acted as meiotic hotspots, but unlike M26 they must be followed by A or C for Atf1.Pcr1 binding and hotspot activity. The basis of the hotspot activity of CTGACGTA (ade6-3013) appears to be identical to that of M26: hotspot activity of both sequences was abolished in cells mutant for atf1, pcr1, spc1, or wis1 and was undetectable in mitotic recombination and in meiotic recombination when located on a plasmid. Both hotspot sequences were sites of micrococcal nuclease hypersensitivity in meiotic chromatin, suggesting that they create an open chromatin structure during meiosis at the site of the hotspots. The newly identified hotspot sequences (C/T/G)TGACGT(A/C) and M26 are closely related to the cAMP response element (CRE) consensus sequence for binding of cAMP-responsive transcription factors such as Atf1.Pcr1, suggesting a link between transcription and meiotic recombination. These results significantly expand the list of identified sequences with meiotic recombination hotspot activity in S. pombe from a single sequence to a family of CRE-related sequences.
七聚体序列ATGACGT对于粟酒裂殖酵母ade6基因中M26减数分裂重组热点的活性至关重要。热点活性与异二聚体转录因子Atf1.Pcr1与M26的结合相关。我们发现序列(C/T/G)TGACGT也能结合Atf1.Pcr1并作为减数分裂热点起作用,但与M26不同的是,它们后面必须跟着A或C才能实现Atf1.Pcr1的结合和热点活性。CTGACGTA(ade6 - 3013)的热点活性基础似乎与M26相同:在atf1、pcr1、spc1或wis1突变的细胞中,这两个序列的热点活性均被消除,并且当位于质粒上时,在有丝分裂重组和减数分裂重组中均检测不到。这两个热点序列都是减数分裂染色质中微球菌核酸酶超敏位点,表明它们在减数分裂期间在热点位点形成开放染色质结构。新鉴定出的热点序列(C/T/G)TGACGT(A/C)和M26与cAMP反应元件(CRE)共有序列密切相关,该共有序列用于结合cAMP反应性转录因子如Atf1.Pcr1,这表明转录与减数分裂重组之间存在联系。这些结果显著地将粟酒裂殖酵母中已鉴定的具有减数分裂重组热点活性的序列列表从单个序列扩展到了一个与CRE相关的序列家族。