Molecular cloning of the calcium and sodium ATPases in Neurospora crassa.
Using PCR, reverse transcription-PCR (RT-PCR) and colony hybridization in a genomic library, we isolated six genes which encode type II P-type ATPases in Neurospora crassa. The six full-length cDNAs were cloned in a yeast expression vector and transformed into Saccharomyces cerevisiae null Ca2+- or Na+-ATPase mutants. Three cDNAs suppressed the defect of the Ca2+ mutant and two of these protected from Mn2+ toxicity. One cDNA suppressed the defect of the Na+ mutant and two cDNAs were not functional in S. cerevisiae. The expression of the transcripts of the six genes in the presence of Ca2+, Na+, high pH or supporting an osmotic shock indicated that, with the exception of one of the Ca2+-ATPases, the main function of the cloned ATPases is the adaptation to stress conditions. The relationship between the cloned fungal Ca2+- and Na+-ATPases and plant type II P-ATPases is discussed.
Benito
B
B
Departamento de Biotecnología, Escuela Técnica Superior de Ingenieros Agrónomos, Universidad Politécnica de Madrid, E-28040 Madrid, Spain.
Garciadeblás
B
B
Rodríguez-Navarro
A
A
D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D027682:Cation Transport Proteins; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000201:enzymology* / Q000378:metabolism; D014176:Protein Biosynthesis; D017386:Sequence Homology, Amino Acid
D027682:Cation Transport Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D000251:Adenosine Triphosphatases; C021838:sodium-translocating ATPase; D000252:Calcium-Transporting ATPases; D002118:Calcium