Allen L A, Morand O H, Raetz C R
Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin, Madison 53706.
Proc Natl Acad Sci U S A. 1989 Sep;86(18):7012-6. doi: 10.1073/pnas.86.18.7012.
Hybrids constructed by fusion of wild-type Chinese hamster ovary cells (CHO-K1) to peroxisome-deficient CHO mutants (ZR-78.1) contain normal peroxisomes, demonstrating that the mutation(s) are recessive. "Nuclear hybrids" prepared by fusion of CHO-K1 karyoplasts to mutant ZR-78.1 occasionally fail to regain intact peroxisomes (approximately 1/300 cells). These peroxisome-deficient nuclear hybrids closely resemble the original mutant cells by biochemical criteria, but their modal chromosome number is 36-38, the same as that of CHO hybrids generated from intact cells. When the peroxisome-deficient nuclear hybrids are fused to wild-type cytoplasts, a fraction of the fusion products (at least 70%) continue to propagate normal peroxisomes indefinitely. Peroxisome biogenesis cannot be reinitiated in cells of mutant ZR-78.1 by fusion to wild-type cytoplasts. Our results suggest that a wild-type nucleus by itself is necessary but not sufficient for restoration of normal peroxisome biogenesis and that a cytoplasmic component of wild-type cells, possibly a normal peroxisome, is also required.
通过将野生型中国仓鼠卵巢细胞(CHO-K1)与过氧化物酶体缺陷的CHO突变体(ZR-78.1)融合构建的杂种细胞含有正常的过氧化物酶体,这表明该突变是隐性的。通过将CHO-K1核体与突变体ZR-78.1融合制备的“核杂种”偶尔无法重新获得完整的过氧化物酶体(约1/300细胞)。这些过氧化物酶体缺陷的核杂种在生化标准上与原始突变细胞非常相似,但其众数染色体数为36 - 38,与由完整细胞产生的CHO杂种相同。当过氧化物酶体缺陷的核杂种与野生型胞质体融合时,一部分融合产物(至少70%)会无限期地继续产生正常的过氧化物酶体。通过与野生型胞质体融合,突变体ZR-78.1细胞中的过氧化物酶体生物发生无法重新启动。我们的结果表明,野生型细胞核本身对于恢复正常的过氧化物酶体生物发生是必要的,但不是充分的,野生型细胞的细胞质成分,可能是正常的过氧化物酶体,也是必需的。