Localization of mitochondrial DNA encoded cytochrome c oxidase subunits I and II in rat pancreatic zymogen granules and pituitary growth hormone granules.
Cytochrome c oxidase (COX) complex is an integral part of the electron transport chain. Three subunits of this complex (COX I, COX II and COX III) are encoded by mitochondrial (mit-) DNA. High-resolution immunogold electron microscopy has been used to study the subcellular localization of COX I and COX II in rat tissue sections, embedded in LR Gold resin, using monoclonal antibodies for these proteins. Immunofluorescence labeling of BS-C-1 monkey kidney cells with these antibodies showed characteristic mitochondrial labeling. In immunogold labeling studies, the COX I and COX II antibodies showed strong and specific mitochondrial labeling in the liver, kidney, heart and pancreas. However, in rat pancreatic acinar tissue, in addition to mitochondrial labeling, strong and specific labeling was also observed in the zymogen granules (ZGs). In the anterior pituitary, strong labeling with these antibodies was seen in the growth hormone secretory granules. In contrast to these compartments, the COX I or COX II antibodies showed only minimal labeling (five- to tenfold lower) of the cytoplasm, endoplasmic reticulum and the nucleus. Strong labeling with the COX I or COX II antibodies was also observed in highly purified ZGs from bovine pancreas. The observed labeling, in all cases, was completely abolished upon omission of the primary antibodies. These results provide evidence that, similar to a number of other recently studied mit-proteins, COX I and COX II are also present outside the mitochondria. The presence of mit-DNA encoded COX I and COX II in extramitochondrial compartments, provides strong evidence that proteins can exit, or are exported, from the mitochondria. Although the mechanisms responsible for protein exit/export remain to be elucidated, these results raise fundamental questions concerning the roles of mitochondria and mitochondrial proteins in diverse cellular processes in different compartments.
Sadacharan
Skanda K
SK
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Canada, L8N 3Z5.
Singh
Bhag
B
Bowes
Timothy
T
Gupta
Radhey S
RS
D000818:Animals; D002417:Cattle; D004272:DNA, Mitochondrial / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics* / Q000378:metabolism; D004792:Enzyme Precursors; D019084:Fluorescent Antibody Technique, Indirect; D013006:Growth Hormone / Q000378:metabolism*; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D046790:Pancreas, Exocrine / Q000201:enzymology* / Q000648:ultrastructure; D010903:Pituitary Gland, Anterior / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D022142:Secretory Vesicles / Q000201:enzymology* / Q000648:ultrastructure
D004272:DNA, Mitochondrial; D004792:Enzyme Precursors; D013006:Growth Hormone; C117108:cytochrome C oxidase subunit II; D003576:Electron Transport Complex IV