Characterization and expression of two genes encoding isoforms of a putative Na, K-ATPase in the chytridiomycete Blastocladiella emersonii.
A P-type ATPase gene (BePAT1) from the aquatic fungus Blastocladiella emersonii, which surprisingly showed high similarity with the alpha-subunit of Na, K-ATPases from animal cells, has been reported recently [Biochim. Biophys. Acta 1383 (1998) 183]. In the present study, we describe the characterization of a second gene, denominated BePAT2, and show that these two genes have a different intron-exon structure but encode putative proteins with greater than 90% amino acid identity. Northern blot and multiplex reverse transcription and polymerase chain reaction (RT-PCR) assays have revealed that BePAT1 and BePAT2 genes have a non-coordinate, developmentally regulated expression during B. emersonii life cycle. Phosphoenzyme formation experiments using the immunopurified enzymes have indicated the presence of a Na, K-ATPase-like activity. Furthermore, immunofluorescence studies using B. emersonii zoospores localized the ATPases on the plasma membrane of these cells.
Fietto
Luciano Gomes
LG
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes, 748-São Paulo-SP 05508-900, Brazil.
Pugliese
Luciana
L
Gomes
Suely Lopes
SL
D001483:Base Sequence; D001754:Blastocladiella / Q000235:genetics*; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification; D019143:Evolution, Molecular; D005800:Genes, Fungal; D007527:Isoenzymes / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D011233:Precipitin Tests; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D013172:Spores, Fungal; D013347:Subcellular Fractions / Q000378:metabolism
D018076:DNA, Complementary; D007527:Isoenzymes; D000254:Sodium-Potassium-Exchanging ATPase