Yeh E, Carbon J, Bloom K
Mol Cell Biol. 1986 Jan;6(1):158-67. doi: 10.1128/mcb.6.1.158-167.1986.
We used DNA fragments from the centromere regions of yeast (Saccharomyces cerevisiae) chromosomes III and XI to examine the transcriptional activity within this chromosomal domain. DNA transcripts were found 200 to 300 base pairs from the 250-base-pair centromere core and lie within an ordered chromatin array. No transcripts were detected from the functional centromere region. We examined the cellular function of one of these tightly centromere-linked transcripts. (CEN11)L, by disrupting the coding sequences in vivo and analyzing the phenotype of the mutant yeast cell. Diploids heterozygous for the (CEN11)L disruption sporulated at wild-type levels, and the absence of the (CEN11)L gene product had no effect on the viability or mitotic growth of haploid cells. Diploids homozygous for the (CEN11)L disruption were unable to sporulate when induced by the appropriate nutritional cues. The mutant cells were competent for intragenic recombination and appeared to be blocked at the mononucleate stage. The temporal ordering of (CEN11)L function with respect to the sporulation mutant spo13 suggests that the (CEN11)L gene product may be required at both the first and second meiotic cell divisions. This new sporulation gene has been termed SPO15.
我们使用来自酵母(酿酒酵母)染色体III和XI着丝粒区域的DNA片段,来检测该染色体区域内的转录活性。在距离250个碱基对的着丝粒核心200到300个碱基对处发现了DNA转录本,它们位于有序的染色质阵列中。在功能着丝粒区域未检测到转录本。我们通过在体内破坏编码序列并分析突变酵母细胞的表型,研究了其中一个与着丝粒紧密相连的转录本(CEN11)L的细胞功能。(CEN11)L破坏杂合的二倍体在野生型水平下产孢,(CEN11)L基因产物的缺失对单倍体细胞的活力或有丝分裂生长没有影响。(CEN11)L破坏纯合的二倍体在受到适当的营养信号诱导时无法产孢。突变细胞能够进行基因内重组,似乎在单核阶段受阻。(CEN11)L功能相对于产孢突变体spo13的时间顺序表明,(CEN11)L基因产物可能在减数分裂的第一次和第二次细胞分裂中都需要。这个新的产孢基因被命名为SPO15。