The yeast mitochondrial transport proteins: new sequences and consensus residues, lack of direct relation between consensus residues and transmembrane helices, expression patterns of the transport protein genes, and protein–protein interactions with other proteins

Abstract

Mitochondrial transport proteins (MTP) typically are homodimeric with a 30-kDa subunit with six transmembrane helices. The subunit possesses a sequence motif highly similar to Pro X Asp/Glu X X Lys/Arg X Arg within each of its three similar 10-kDa segments. Four (YNL083W, YFR045W, YPR021C, YDR470C) of the 35 yeast (S. cerevisiae) MTP genes were resequenced since the masses of their proteins deviate significantly from the typical 30 kDa. We now find these four proteins to have 545, 285, 902, and 502 residues, respectively. Together with only four other MTPs, the sequences of YPR021C and YDR470C show substitutions of some of the five residues that are absolutely conserved among the 12 MTPs with identified transport function and 17 other MTPs. We do now find these five consensus residues also in the new sequences of YNL083W and YFR045W. Additional analyses of the 35 yeast MTPs show that the location of transmembrane helix sequences do not correlate with the general consensus residues of the MTP family; protein segments connecting the six transmembrane helices and facing the intermembrane space are not uniformly short (about 20 residues) or long (about 40 residues) when facing the matrix; most MTPs have at least one transmembrane helix for which the sum of the negative hydropathy values of all residues yields a very small negative value, suggesting a membrane location bordering polar faces of other transmembrane helices or a non-transmembrane location. The extra residues of the three large MTPs are hydrophilic and at the N-terminal. The 200-residue N-terminal segment of YNL083W has four putative Ca2+-binding sites. The 500-residue N-terminal segment of YPR021C shows sequence similarity to enzymes of nucleic acid metabolism. cDNA microarray data show that YNL083W is expressed solely during sporulation, while the expressions of YFR045W, YPR021C, and YDR470C are induced by various stress situations. These results also show that the 35 MTP genes are expressed under a rather diverse set of metabolic conditions that may help identify the function of the proteins. Interestingly, yeast two-hybrid screens, that will also be useful in identifying the function of MTPs, indicate that MIR1, AAC3, YOR100C, and YPR011C do interact with non-MTPs.

Polymorphisms in Multiple Genes Contribute to the Spontaneous Mitochondrial Genome Instability ofSaccharomyces cerevisiaeS288C Strains

“…SAL1 contains a frameshift mutation (the sal1-1 allele) in certain laboratory strains (Chen 2004). The locus history page in the SGD for SAL1 documents that the sal1-1 allele was corrected in the database in February 2004 after sequence comparisons to related fungi (Belenkiy et al 2000;Brachat et al 2003). We sequenced the SAL1 gene from BY4716 and S288C.…”

Plant Colonization by the Vascular Wilt Fungus Fusarium oxysporum Requires FOW1, a Gene Encoding a Mitochondrial Protein

“…B60 showed markedly reduced virulence to melon plants . The affected gene, named FOW1 , encodes a protein with high similarity to mitochondrial carrier proteins (MCPs) of yeast (Nelson et al, 1998;Belenkiy et al, 2000). MCPs are small transport proteins of the mitochondrial inner membrane that catalyze the transport of metabolites across the inner membrane with a high degree of substrate specificity (Palmieri, 1994;Nelson et al, 1998;Belenkiy et al, 2000).…”

Deletion of the Mitochondrial Carrier Genes MRS3 andMRS4 Suppresses Mitochondrial Iron Accumulation in a Yeast Frataxin-deficient Strain

“…Saccharomyces cerevisiae contains 35 of these carriers (15,16), the most studied of which is the mitochondrial ATP/ADP translocator. Mitochondrial carriers are characterized by three homologous domains, each containing two transmembrane spans.…”