Dutcher S K
Department of Genetics, University of Washington, Seattle 98195.
Mol Cell Biol. 1981 Mar;1(3):245-53. doi: 10.1128/mcb.1.3.245-253.1981.
Heterokaryons of Saccharomyces cerevisiae have been constructed utilizing the kar1-1 mutation, which prevents nuclear fusion during conjugation (J. Conde and G. Fink, Proc. Natl. Acad. Sci. U.S.A. 73:3651-3655, 1976). Each heterokaryon contained two haploid nuclei that were marked on several chromosomes. They segregated haploid progeny (cytoductants), most of which have the nuclear genotype of one or the other of the heterokaryon parents, but they occasionally segregated progeny having a recombinant genotype (exceptional cytoductants). Exceptional cytoductants receive the majority of their genome from one parent (the recipient) and a minority from the other (the donor). Transfer of two markers from the donor nucleus to the recipient is rarely coincident for markers located on different chromosomes but is nearly always coincident for those markers located on the same chromosome, suggesting that whole chromosomes are transferred from the donor nucleus to the recipient. In crosses of kar1-1 X KAR1 parents, either nucleus may act as a recipient or donor with equal probability. Recipient nuclei acquired 9 of the 10 chromosomes examined, with frequencies which were inversely correlated with the size of the chromosome. When a chromosome is acquired by the recipient nucleus, it either replaces its homolog or exists in a disomic condition. Haploid progeny emanating from kar1 X KAR1 crosses are frequently inviable. I tested whether this inviability might be the result of chromosome loss by donor nuclei. Viability of progeny from kar1 X KAR1 heterokaryons was improved when the parental nuclei were diploid to an extent consistent with the hypothesis, and diploid progeny which had become monosomic were recovered from these heterokaryons. The following sequence of events accounts for chromosome transfer in kar1 X KAR1 heterokaryons. After cell fusion, each nucleus in the heterokaryon has a probability of about 0.38 of losing one or more chromosomes. A nucleus sustaining such a loss can become a donor in a chromosome transfer event. If the other nucleus does not sustain a mortal chromosome loss, it can become a recipient in a transfer event. The chance of acquiring a chromosome lost by the donor is greater for smaller chromosomes than for larger ones and is about 0.05 for the average chromosome.
利用kar1-1突变构建了酿酒酵母的异核体,该突变可阻止接合过程中的核融合(J. 康德和G. 芬克,《美国国家科学院院刊》73:3651 - 3655,1976年)。每个异核体包含两个单倍体核,这两个核在几条染色体上都有标记。它们分离出单倍体后代(胞质融合子),其中大多数具有异核体亲本一方或另一方的核基因型,但偶尔也会分离出具有重组基因型的后代(异常胞质融合子)。异常胞质融合子从一个亲本(受体)获得其大部分基因组,从另一个亲本(供体)获得少数基因组。对于位于不同染色体上的标记,供体核中两个标记转移到受体几乎不会同时发生,但对于位于同一染色体上的那些标记,几乎总是同时发生,这表明整条染色体从供体核转移到受体。在kar1-1×KAR1亲本的杂交中,任何一个核作为受体或供体的概率相等。受体核获得了所检测的10条染色体中的9条,其频率与染色体大小呈负相关。当受体核获得一条染色体时,它要么取代其同源染色体,要么以二体状态存在。kar1×KAR1杂交产生的单倍体后代通常无法存活。我测试了这种无法存活是否可能是供体核染色体丢失的结果。当亲本核为二倍体时,kar1×KAR1异核体后代的活力有所提高,这在一定程度上与该假设相符,并且从这些异核体中回收了已成为单体的二倍体后代。以下事件序列解释了kar1×KAR1异核体中的染色体转移。细胞融合后,异核体中的每个核有大约0.38的概率丢失一条或多条染色体。遭受这种丢失的核可以在染色体转移事件中成为供体。如果另一个核没有遭受致命的染色体丢失,它可以在转移事件中成为受体。较小染色体获得供体丢失染色体的机会比较大染色体更大,平均每条染色体的机会约为0.05。