Raymond W E, Kleckner N
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Mol Gen Genet. 1993 Apr;238(3):390-400. doi: 10.1007/BF00291998.
In Saccharomyces cerevisiae, the RAD50 gene is required for repair of X-ray and MMS-induced DNA damage during vegetative growth, and for synaptonemal complex formation and genetic recombination during meiosis. We show below that the RAD50 gene encodes major and minor transcripts of 4.2 and 4.6 kb in length which differ primarily at their 5' ends. Steady-state levels of both RAD50 transcripts increase coordinately during meiosis, reaching maximal levels midway through meiotic prophase, about 3 or 4 h after transfer of cells to sporulation medium. The 5' ends of the major RAD50 transcript in both meiotic and vegetative cells map to the same cluster of sites approximately 20 bp upstream of the amino-terminal ATG of the RAD50 coding sequence. We conclude that the increased RAD50 transcript level observed during meiosis does not reflect utilization of a new promoter. In contrast, steady-state levels of Rad50 protein do not increase during meiosis. Thus, changes in RAD50 transcript levels are not necessarily accompanied by commensurate changes in Rad50 protein levels. Possible explanations are considered.
在酿酒酵母中,RAD50基因在营养生长期间对X射线和甲基磺酸甲酯(MMS)诱导的DNA损伤修复是必需的,在减数分裂期间对联会复合体形成和基因重组也是必需的。我们在下文表明,RAD50基因编码长度为4.2 kb和4.6 kb的主要和次要转录本,它们主要在5'端存在差异。在减数分裂期间,两种RAD50转录本的稳态水平协同增加,在减数分裂前期中期达到最高水平,即细胞转移到产孢培养基后约3或4小时。减数分裂和营养细胞中主要RAD50转录本的5'端都定位到RAD50编码序列氨基端ATG上游约20 bp处的同一簇位点。我们得出结论,减数分裂期间观察到的RAD50转录本水平增加并不反映新启动子的利用。相反,减数分裂期间Rad50蛋白的稳态水平没有增加。因此,RAD50转录本水平的变化不一定伴随着Rad50蛋白水平的相应变化。文中考虑了可能的解释。