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

Mitochondrial transport proteins (MTP) typically are homodimeric with a 30- kDa subunit with six transmembrane helices. The subunit possesses a sequenc e motif highly similar to Pro X Asp/Glu X X Lys/Arg X Arg within each of it s 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 fi nd these four proteins to have 545, 285, 902, and 502 residues, respectivel y. Together with only four other MTPs, the sequences of YPR021C and YDR470C show substitutions of some of the five residues that are absolutely conser ved 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 genera l 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 matr ix; most MTPs have at least one transmembrane helix for which the sum of th e negative hydropathy values of all residues yields a very small negative v alue, suggesting a membrane location bordering polar faces of other transme mbrane helices or a non-transmembrane location. The extra residues of the t hree large MTPs are hydrophilic and at the N-terminal. The 200-residue N-te rminal segment of YNL083W has four putative Ca2+-binding sites. The 500-res idue N-terminal segment of YPR021C shows sequence similarity to enzymes of nucleic acid metabolism. cDNA microarray data show that YNL083W is expresse d 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. Interestin gly, yeast two-hybrid screens, that will also be useful in identifying the function of MTPs, indicate that MIR1, AAC3, YOR100C, and YPR011C do interac t with non-MTPs. (C) 2000 Elsevier Science B.V. All rights reserved.