Molecular cloning of the calcium and sodium ATPases in Neurospora crassa

Mol Microbiol. 2000 Mar;35(5):1079-88. doi: 10.1046/j.1365-2958.2000.01776.x.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / chemistry
  • Adenosine Triphosphatases / genetics*
  • Adenosine Triphosphatases / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Calcium / metabolism
  • Calcium-Transporting ATPases / chemistry
  • Calcium-Transporting ATPases / genetics*
  • Calcium-Transporting ATPases / metabolism
  • Cation Transport Proteins*
  • Cloning, Molecular
  • DNA Primers
  • DNA, Complementary
  • Molecular Sequence Data
  • Neurospora crassa / enzymology*
  • Neurospora crassa / metabolism
  • Protein Biosynthesis
  • Sequence Homology, Amino Acid

Substances

  • Cation Transport Proteins
  • DNA Primers
  • DNA, Complementary
  • Adenosine Triphosphatases
  • sodium-translocating ATPase
  • Calcium-Transporting ATPases
  • Calcium

Associated data

  • GENBANK/AJ243515
  • GENBANK/AJ243516
  • GENBANK/AJ243517
  • GENBANK/AJ243518
  • GENBANK/AJ243519
  • GENBANK/AJ243520