The correlation between the import of the Rieske iron-sulfur protein into the mitochondrial matrix and processing of the precursor protein by matrix processing peptidase was investigated using high concentrations of metal chelators and iron-sulfur protein in which the recognition site for the matrix processing peptidase was destroyed by site-directed mutagenesis. High concentrations of EDTA and...

Despite the wealth of knowledge regarding the mechanisms of action and the mechanisms of resistance to azole antifungals, very little is known about how the azoles are imported into pathogenic fungal cells. Here the in-vitro accumulation and import of Fluconazole (FLC) was examined in the pathogenic fungus, Candida albicans. In energized cells, FLC accumulation correlates inversely with express...

Nuclear import of plasmid DNA in nondividing cells is a process essential to the success of numerous viral life cycles, gene therapy protocols, and gene expression experiments. Here, intact protein-free SV40 DNA was cytoplasmically injected into cells and its subcellular localization was followed by in situ hybridization. SV40 DNA localized to the nucleus consistent with a mechanism of transpor...

Most mitochondrial proteins are imported into mitochondria through transmembrane channels composed largely, and perhaps exclusively, of proteins. We have determined the effective internal diameter of the protein import channel in the mitochondrial outer membrane to be between 20 A and 26 A during translocation. The diameter of the import channel in the inner membrane is smaller than the diamete...

Mitochondria import a large number of nuclear-encoded proteins via membrane-bound transport machineries; however, little is known about regulation of the preprotein translocases. We report that the main protein entry gate of mitochondria, the translocase of the outer membrane (TOM complex), is phosphorylated by cytosolic kinases-in particular, casein kinase 2 (CK2) and protein kinase A (PKA). C...

We have transcribed mRNA from a cDNA clone coding for pea ribulose-1,5-bisphosphate carboxylase, translated the mRNA in a wheat germ cell-free system, and studied the energy requirement for posttranslational import of the [35S]methionine-labeled protein into the stroma of pea chloroplasts. We found that import depends on ATP hydrolysis within the stroma. Import is not inhibited when H+, K+, Na+...

Chloroplasts must import a large number of proteins from the cytosol. It generally is assumed that this import proceeds for all stromal and thylakoid proteins in an identical manner and is caused by the operation of two distinctive protein import machineries in the outer and inner plastid envelope, which form the general import site. Here we show that there is a second site of protein transloca...

To identify new components that mediate mitochondrial protein import, we analyzed mas6, an import mutant in the yeast Saccharomyces cerevisiae. mas6 mutants are temperature sensitive for viability, and accumulate mitochondrial precursor proteins at the restrictive temperature. We show that mas6 does not correspond to any of the presently identified import mutants, and we find that mitochondria ...

SRY and other Sox-type transcription factors are important developmental regulators with various implications in human disease. In this study, we identified Exp4 (exportin 4) as an interaction partner of Sox2 in mouse embryonic stem cells and neural progenitors. We show that, besides its established function in nuclear export, Exp4 acts as a bona fide nuclear import receptor for Sox2 and SRY. T...