Scott J M, Mikami K, Leeck C L, Forney J D
Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907-1153.
Mol Cell Biol. 1994 Apr;14(4):2479-84. doi: 10.1128/mcb.14.4.2479-2484.1994.
Paramecium tetraurelia contains two types of nuclei, a diploid germinal micronucleus and a large transcriptionally active macronucleus. The macronuclear genome is formed from the micronuclear DNA during sexual reproduction. Previous studies have shown that the processing of the A-type variable surface protein gene during formation of a new macronucleus is dependent on the presence of the A gene in the old macronucleus. It is not clear if this is a general feature that controls the formation of the Paramecium macronuclear genome or a unique feature of the A locus. Using micronuclear transplantation, we have constructed a strain that has a wild-type micronucleus but has macronuclear deletions of the A- and B-type surface protein genes. Neither the A nor the B gene is incorporated into the new macronucleus after sexual reproduction. Macronuclear transformation of this strain with the B gene rescues the B-gene deletion after formation of the next macronucleus but has not effect on the A deletion. Similarly, transformation with the A gene shows gene-specific rescue for A but not B. The effect of the old macronucleus on the processing of the new macronucleus results in a pattern of non-Mendelian inheritance of both macronuclear deletions. Progeny from the wild-type exconjugant are all wild type, and progeny from the A- B- exconjugant are mutant. The features of this A- B- non-Mendelian mutant demonstrate that the regulation of macronuclear DNA processing is gene specific, and our results open the possibility that this type of regulation affects many regions of the Paramecium genome.
四膜虫含有两种类型的细胞核,一种是二倍体生殖性小核,另一种是大型转录活性大核。大核基因组在有性生殖过程中由小核DNA形成。先前的研究表明,新大核形成过程中A型可变表面蛋白基因的加工依赖于旧大核中A基因的存在。目前尚不清楚这是控制四膜虫大核基因组形成的普遍特征,还是A基因座的独特特征。通过微核移植,我们构建了一个菌株,它具有野生型小核,但大核中A型和B型表面蛋白基因缺失。有性生殖后,A基因和B基因都不会整合到新的大核中。用B基因对该菌株进行大核转化,可在下一个大核形成后挽救B基因缺失,但对A基因缺失没有影响。同样,用A基因转化对A基因有特异性挽救作用,但对B基因没有。旧大核对新大核加工的影响导致了两种大核缺失的非孟德尔遗传模式。野生型接合后体的后代都是野生型,而A - B -接合后体的后代是突变型。这种A - B -非孟德尔突变体的特征表明,大核DNA加工的调控是基因特异性的,我们的结果开启了这种调控类型影响四膜虫基因组许多区域的可能性。