Phe356 in the yeast Ca2+ channel component Mid1 is a key residue for viability after exposure to alpha-factor.
The yeast Mid1 protein is an integral membrane protein required for the viability of differentiated cells and Ca2+ influx induced by mating pheromone. Our previous study has identified a loss-of-function mutation, F356S. The F356S mutant is completely unable to maintain viability, but still has Ca2+ accumulation activity near the wild-type level. Here we further examined in detail the F356S mutation to unravel the function of Phe356. After exposure to the pheromone, the F356S mutant was not fully rescued by high extracellular Ca2+, like the mid1 null mutant, suggesting that Phe356 and Mid1 itself are also required for viability maintenance mechanism that does not involve Ca2+ signalling. Substitutions of hydrophilic amino acids for Phe356 caused lethality and low Ca2+ accumulation, but those of hydrophobic amino acids did not. Substitutions of small amino acids for Phe356 caused a significantly reduced viability, but did not affect Ca2+ accumulation. We suggest that the hydrophobicity of the Phe356 residue is important for both viability maintenance and Ca2+ uptake, and that its size for viability maintenance.
Tada
Tomoko
T
Department of Biology, Tokyo Gakugei University, 4-1-1 Nukuikita-machi, Koganei-shi, Tokyo 184-8501, Japan.
Ohmori
Masayuki
M
Iida
Hidetoshi
H
D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000737:chemistry*; D002122:Calcium Chloride / Q000494:pharmacology; D002470:Cell Survival; D003545:Cysteine / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010641:Phenotype; D010649:Phenylalanine / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000502:physiology*; D015398:Signal Transduction; D013997:Time Factors
D015220:Calcium Channels; D005656:Fungal Proteins; C090463:MID1 protein, S cerevisiae; D008562:Membrane Glycoproteins; D029701:Saccharomyces cerevisiae Proteins; D010649:Phenylalanine; D003545:Cysteine; D002122:Calcium Chloride; D002118:Calcium