Moens P B, Freire R, Tarsounas M, Spyropoulos B, Jackson S P
Department of Biology, York University, Toronto, Ontario, M3J 1P3, Canada.
J Cell Sci. 2000 Feb;113 ( Pt 4):663-72. doi: 10.1242/jcs.113.4.663.
Bloom's syndrome (BS) is a recessive human genetic disorder characterized by short stature, immunodeficiency and elevated risk of malignancy. BS cells have genomic instability and an increased frequency of sister chromatid exchange. The gene mutated in BS, BLM, encodes a 3'-5' helicase (BLM) with homology to bacterial recombination factor, RecQ. Human males homozygous for BLM mutations are infertile and heterozygous individuals display increased frequencies of structural chromosome abnormalities in their spermatozoa. Also, mutations in the Saccharomyces cerevisiae homolog of BLM, Sgs1, cause a delay in meiotic nuclear division and a reduction in spore viability. These observations suggest that BLM may play a role during meiosis. Our antibodies raised against the C terminus of the human protein specifically recognize both mouse and human BLM in western blots of cell lines and in successive developmental stages of spermatocytes, but fail to detect BLM protein in a cell line with a C-terminally truncated protein. BLM protein expression and location are detected by immunofluorescence and immunoelectron microscopy as discrete foci that are sparsely present on early meiotic prophase chromosome cores, later found abundantly on synapsed cores, frequently in combination with the recombinases RAD51 and DMC1, and eventually as pure BLM foci. The colocalization of RAD51/DMC1 with BLM and the statistically significant excess of BLM signals in the synapsed pseudoautosomal region of the X-Y chromosomes, which is a recombinational hot spot, provide indications that BLM protein may function in the meiotic recombination process.
布卢姆综合征(BS)是一种隐性人类遗传病,其特征为身材矮小、免疫缺陷以及患恶性肿瘤的风险增加。BS细胞具有基因组不稳定性,姐妹染色单体交换频率增加。在BS中发生突变的基因BLM编码一种3'-5'解旋酶(BLM),与细菌重组因子RecQ具有同源性。BLM突变的纯合人类男性不育,杂合个体精子中结构染色体异常的频率增加。此外,BLM在酿酒酵母中的同源物Sgs1发生突变会导致减数分裂核分裂延迟以及孢子活力降低。这些观察结果表明BLM可能在减数分裂过程中发挥作用。我们针对人类蛋白质C末端产生的抗体在细胞系的蛋白质印迹以及精母细胞的连续发育阶段中能特异性识别小鼠和人类的BLM,但在一个C末端截短蛋白的细胞系中未能检测到BLM蛋白。通过免疫荧光和免疫电子显微镜检测到BLM蛋白的表达和定位为离散的病灶,这些病灶在减数分裂前期早期染色体核心上稀疏存在,后来在联会的核心上大量出现,经常与重组酶RAD51和DMC1结合,最终成为纯BLM病灶。RAD51/DMC1与BLM的共定位以及在X-Y染色体的联会假常染色体区域(一个重组热点)中BLM信号在统计学上的显著过量,表明BLM蛋白可能在减数分裂重组过程中发挥作用。