ETIOLATED CELLS OF CHLAMYDOMONAS-REINHARDTII - CHOICE MATERIAL FOR CHARACTERIZATION OF MITOCHONDRIAL-MEMBRANE POLYPEPTIDES

被引:11
作者
BENNOUN, P
ATTEIA, A
PIERRE, Y
DELOSME, M
机构
[1] Inst. de Biologie Physico-Chimique, Ctr. Natl. de la Rech. Scientifique, Unité Associée 1187, 75005 Paris, 13, rue Pierre et Marie Curie
关键词
D O I
10.1073/pnas.92.22.10202
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We have investigated a light-conditional mutant of Chlamydomonas reinhardtii (J12) that is unable to synthesize chlorophyll in the dark with the aim of characterizing the mitochondrial membrane polypeptides of this alga. A crude membrane fraction derived from etiolated cells was analyzed by gel electrophoresis, immunoblot analysis, and pulse-labeling in the presence of specific protein synthesis inhibitors, This fraction contained both mitochondrial and etioplast membranes, and the latter contained appreciable amounts of subunits of the cytochrome b(6)f complex. The mitochondria-encoded subunit 1 of cytochrome-c oxidase called COX1 was identified, and its synthesis was detected in this membrane fraction. The redox-difference spectra of mitochondrial cytochromes were studied in whole cells and membrane fractions, in both respiratory-competent and -deficient strains. Mitochondrial membranes could be further purified after sucrose gradient centrifugation, The use of etiolated cells and their membrane extracts, in association with appropriate methodologies, opens ways to study the molecular genetics of mitochondria in C. reinhardtii and allows us to address the question of the cooperation established between the three genetic compartments of a plant cell.
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页码:10202 / 10206
页数:5
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