Molecular cloning of the calcium and sodium ATPases in Neurospora crassa

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Abstract

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.

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Benito, B., Garciadeblás, B., & Rodríguez-Navarro, A. (2000). Molecular cloning of the calcium and sodium ATPases in Neurospora crassa. Molecular Microbiology, 35(5), 1079–1088. https://doi.org/10.1046/j.1365-2958.2000.01776.x

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