Significant effect of the N-terminal region of the mitochondrial ADP/ATP carrier on its efficient expression in yeast mitochondria.
The low-level expression of the bovine heart mitochondrial ADP/ATP carrier (bovine type 1 ADP/ATP carrier (bAAC1)) in the yeast mitochondrial membrane is significantly improved by replacement of its N-terminal region with corresponding regions of the yeast type 1 and 2 carriers (yAAC1 and yAAC2) (Hashimoto, M., Shinohara, Y., Majima, E., Hatanaka, T., Yamazaki, N., and Terada, H. (1999) Biochim. Biophys. Acta 1409, 113--124). To understand why the bAAC1 chimeras were highly expressed in yeast mitochondria, we examined the effects of the length and sequence of the N-terminal region extending into the cytosol on the expression of bAAC1 and yAAC2 derivatives in yeast mitochondria. For this, their N-terminal regions were replaced with peptide fragments of various lengths and sequences derived from those of bAAC1, yAAC1, and yAAC2. We found that a specific amino acid sequence and a definite length of the N-terminal region of yAAC2 were required for high expression of bAAC1 and yAAC2 in yeast mitochondria. We also examined the steady-state transcript levels and expression of these derivatives in whole yeast cells. Based on our results, we discuss the role of the N-terminal region in efficient expression of bAAC1 and yAAC2 in yeast mitochondria.
Hatanaka
T
T
Faculty of Pharmaceutical Sciences, University of Tokushima, Shomachi-1, Tokushima 770-8505, Japan.
Hashimoto
M
M
Majima
E
E
Shinohara
Y
Y
Terada
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D017931:DNA Primers; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology*; D000226:Mitochondrial ADP, ATP Translocases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D017931:DNA Primers; C112225:PET9 protein, S cerevisiae; D010446:Peptide Fragments; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D000226:Mitochondrial ADP, ATP Translocases