A PMR1-like calcium ATPase of Aspergillus fumigatus: cloning, identification and functional expression in S. cerevisiae.
The understanding of the controlling factors of calcium homeostasis in Aspergillus fumigatus is very poor, although this ion is involved in several important events of these particular cells. We have cloned, identified and expressed for functional complementation a PMR1-like Ca(2+)-ATPase gene from A. fumigatus. The Afpmr1 gene encodes a protein of 1061 deduced amino acids, containing all the conserved subdomains found in other P-type ATPases: the phosphatase region, phosphorylation site, FITC labelling site, ATP binding domain; E(386), N871, D875 amino acid residues for calcium ion interaction and Q880, a residue that alters ion selectivity in PMR1. The expressed AfPMR1 in S. cerevisiae K616 strain functionally complemented the deficient growth in EGTA (5-20 mM)- and MnCl2 (4 mM)-containing medium. These results demonstrate the first evidence of a Ca(2+)-ATPase in A. fumigatus and strongly suggest a role for this enzyme in calcium and manganese homeostasis.
Soriani
F M
FM
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Ribeirão Preto, São Paulo 14040-903, Brazil.
Martins
V P
VP
Magnani
T
T
Tudella
V G
VG
Curti
C
C
Uyemura
S A
SA
D000595:Amino Acid Sequence; D001232:Aspergillus fumigatus / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000235:genetics*; D002712:Chlorides; D003470:Culture Media; D004533:Egtazic Acid; D005816:Genetic Complementation Test; D017895:Manganese Compounds; D018832:Molecular Chaperones / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
D002712:Chlorides; D003470:Culture Media; D017895:Manganese Compounds; D018832:Molecular Chaperones; C101767:SSC1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D004533:Egtazic Acid; D000252:Calcium-Transporting ATPases; C025340:manganese chloride