Candida glabrata metallothioneins. Cloning and sequence of the genes and characterization of proteins.
Southern blot analysis has identified several metallothionein gene sequences in a human pathogenic yeast Candida glabrata. Two of these genes encoding proteins designated MT-I and MT-II have been cloned and sequenced. No introns were found in either of the genes. The complete primary structure of MT-II was also determined by protein sequencing methods. As isolated, MT-I and MT-II consist of 62 and 51 amino acids, respectively. The only residues predicted from the nucleotide sequence but not present in the isolated protein are the amino-terminal methionines in each sequence. MT-I contains 18 cysteines, 14 of which are present as Cys-X-Cys motifs and two additional cysteines in a Cys-X-X-Cys sequence. The sequence of MT-II contains 16 cysteinyl residues, 14 of which are in Cys-X-Cys sequences. Fluorescence spectroscopy indicates the presence of Cu(I)-thiolate bonds in both proteins. The binding stoichiometries are 11-12 for MT-I and 10 for MT-II. Under certain nutritional conditions, a truncated form of MT-II was also produced. Northern analysis of the total cellular RNA from copper-treated cells showed that both MT-I and MT-II genes are regulated by this metal ion in a concentration-dependent fashion. The concentrations of MT-II mRNA appeared to be higher than that of MT-I mRNA at all concentrations of copper sulfate tested. Both genes are inducible by silver but not by cadmium salts. Cadmium ions, however, are effective in reducing the control levels of both MT-I and MT-II mRNAs.
Mehra
R K
RK
University of Utah Medical Center, Salt Lake City 84132.
Garey
J R
JR
Butt
T R
TR
Gray
W R
WR
Winge
D R
DR
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002175:Candida / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D005800:Genes, Fungal; D008668:Metallothionein / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D015183:Restriction Mapping
D004271:DNA, Fungal; D008668:Metallothionein