Morphology and protein composition of the mitochondrial nucleoids in yeast cells lacking Abf2p, a high mobility group protein
Morphology and protein composition of the mitochondrial nucleoids in yeast cells lacking Abf2p, a high mobility group protein
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DOI：
10.2323/jgam.56.455
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发表时间：
2010-12-01
影响因子：
1.2
通讯作者：
Sato, Hiroshi
中科院分区：
文献类型：
--
作者：
Miyakawa, Isamu;Kanayama, Miwako;Sato, Hiroshi
为了阐明Abf 2 p的作用，一个主要的线粒体！DNA结合蛋白在酿酒酵母中的表达，我们用4 '，6-二脒基-2-苯基吲哚（DAPI）染色法在体内和体外研究了ABF 2缺陷突变体（Δ abf 2）的线粒体类核（mt-nucleoids）的形态。在含有甘油的YPG培养基中生长至对数生长期的Delta abf 2细胞中，mt-类核出现为具有不规则大小的弥散结构，与野生型细胞中mt-类核的串珠外观相反。此外，在对数期，Delta abf 2细胞中传输到芽的mt-类核的DAPI荧光强度显著低于野生型细胞。然而，Abf 2 p的缺乏并不影响线粒体的形态或分离。从在YPG培养基中生长至稳定期的Delta abf 2细胞中分离的mt-类核蛋白的蛋白组成与从在相同条件下培养的野生型细胞中分离的mt-类核蛋白的蛋白组成非常相似，除了缺乏Abf 2 p。这些结果共同表明，在对数期细胞中，Abf 2 p的缺乏不仅影响mt-类核的形态，而且影响它们向芽中的传递。另一方面，我们的研究结果表明，在静止期细胞中，Abf 2 p的缺乏不会显着改变mt-类核蛋白的蛋白质组成。
To elucidate the role of Abf2p, a major mitochondria! DNA-binding protein in the yeast Saccharomyces cerevisiae, we examined the morphology of the mitochondrial nucleoids (mt-nucleoids) in an ABF2-deficient mutant (Delta abf2) in vivo and in vitro by 4',6-diamidino-2-phenylindole (DAPI) staining. The mt-nucleoids appeared as diffuse structures with irregular-size in Delta abf2 cells that were grown to log phase in YPG medium containing glycerol, in contrast to the strings-of-beads appearance of mt-nucleoids in wild-type cells. In addition, DAPI-fluorescence intensity of the mt-nucleoids transmitted to the bud was significantly lower in Delta abf2 cells than in wild-type cells at log phase. However, the lack of Abf2p did not affect the morphology or segregation of mitochondria. The protein composition of the mt-nucleoids isolated from Delta abf2 cells grown to stationary phase in YPG medium was very similar to that of the mt-nucleoids isolated from wild-type cells cultured under the same conditions, except for the lack of Abf2p. These results together suggested that in log-phase cells, the lack of Abf2p influences not only the morphology of mt-nucleoids but also their transmission into the bud. On the other hand, our result suggested that in stationary-phase cells, the lack of Abf2p does not significantly alter the protein composition of the mt-nucleoids.