Development of Mitochondrial Membranes in Anaerobically Grown Yeast Cells1

Abstract

Biochemical analyses of mitochondrial marker substances, especially cardiolipin and oligomycin-sensitive ATPase [EC 3.6.1.3], as well as electron microscopic observations were carried out to eludicate the process of mitochondrial development in annaerobic yeast cells. Cardiolipin was found to be localized in the mitochondria in anaerobic cells. Its cellular content was a little higher in the stationary phase than in the exponential phase in glucose-grown cells and increased further in galactose-grown cells. The lipid content of the mitochondrial preparation obtained from glucose-grown stationary cells was nearly as high as that from galactose-grown cells. It was also comparable to that of aerobic cells in the stationary phase, where mitochondria are fully developed. Both cellular and mitochondrial levels of oligomycin-sensitive ATPase activity were also found to rise markedly in galactose-grown anaerobic cells, although not in stationary phase cells grown anaerobically on glucose. These high levels of the mitochondrial markers indicate a developmental change in mitochondrial structure even in anaerobically grown cells, which lack mitochondrial cytochromes. In the process of aerobic adaptation, respiratory system formation was observed to occur much faster in galactose-grown cells than in glucose-grown cells, and not to be inhibited by chloramphenicol and high concentrations of glucose structure in anaerobic cells. The developmental change was also corroborated by electron microscopic observations, which revealed the occurrence of two types of mitochondria in anaerobic cells. One was found in glucose-repressed cells and was characterized by the presence of numerous electron-dense granules in the matrix. In contrast, the other type, found in glucose-derepressed cells, had an electron-lucent matrix. No crista membrane was seen in either type of mitochondria in anaerobic cells, although the infoldings of the inner membrane, which partition the matrix into two parts and therefore are called "septum membranes," appeared frequently in the stationary phase cells. On the basis of these results, the process of mitochondrial development in yeast cells is discussed.