Surosky R T, Tye B K
Genetics. 1985 Jul;110(3):397-419. doi: 10.1093/genetics/110.3.397.
We have integrated a plasmid containing a yeast centromere, CEN5, into the HIS4 region of chromosome III by transformation. Of the three transformant colonies examined, none contained a dicentric chromosome, but all contained a rearranged chromosome III. In one transformant, rearrangement occurred by homologous recombination between two Ty elements; one on the left arm and the other on the right arm of chromosome III. This event produced a ring chromosome (ring chromosome III) of about 60 kb consisting of CEN3 and all other sequences between the two Ty elements. In addition, a linear chromosome (chromosome IIIA) consisting of sequences distal to the two Ty elements including CEN5, but lacking 60 kb of sequences from the centromeric region, was produced. Two other transformants also contain a similarly altered linear chromosome III as well as an apparently normal copy of chromosome III. These results suggest that dicentric chromosomes cannot be maintained in yeast and that dicentric structures must be resolved for the cell to survive.--The meiotic segregation properties of ring chromosome III and linear chromosome IIIA were examined in diploid cells which also contained a normal chromosome III. Chromosome IIIA and normal chromosome III disjoined normally, indicating that homology or parallel location of the centromeric regions of these chromosomes are not essential for proper meiotic segregation. In contrast, the 60-kb ring chromosome III, which is homologous to the centromeric region of the normal chromosome III, did not appear to pair with fidelity with chromosome III.
我们通过转化将一个含有酵母着丝粒CEN5的质粒整合到了第三条染色体的HIS4区域。在所检测的三个转化菌落中,没有一个含有双着丝粒染色体,但都含有一条重排的第三条染色体。在一个转化体中,重排是由两个Ty元件之间的同源重组发生的;一个在第三条染色体的左臂,另一个在右臂。这一事件产生了一个约60 kb的环状染色体(环状第三条染色体),它由CEN3和两个Ty元件之间的所有其他序列组成。此外,还产生了一个线性染色体(第三条染色体A),它由包括CEN5在内的两个Ty元件远端的序列组成,但缺少着丝粒区域的60 kb序列。另外两个转化体也含有一条类似改变的线性第三条染色体以及一条明显正常的第三条染色体拷贝。这些结果表明,双着丝粒染色体在酵母中无法维持,并且为了细胞存活,双着丝粒结构必须得到解决。——在还含有一条正常第三条染色体的二倍体细胞中检测了环状第三条染色体和线性第三条染色体A的减数分裂分离特性。第三条染色体A和正常第三条染色体正常分离,这表明这些染色体着丝粒区域的同源性或平行位置对于正确的减数分裂分离并非必需。相反,与正常第三条染色体着丝粒区域同源的60 kb环状第三条染色体似乎没有与第三条染色体精确配对。