Functional analysis of the Neurospora crassa PZL-1 protein phosphatase by expression in budding and fission yeast

Yeast. 2001 Jan 30;18(2):115-24. doi: 10.1002/1097-0061(20010130)18:2<115::AID-YEA653>3.0.CO;2-G.

Abstract

The gene pzl-1 from the filamentous fungus Neurospora crassa encodes a putative Ser/Thr protein phosphatase that is reminiscent of the Ppz1/Ppz2 and Pzh1 phosphatases from Saccharomyces cerevisiae and Schizosaccharomyces pombe, respectively. The entire PZL-1 protein, as well as its carboxyl-terminal domain, have been expressed in Escherichia coli as active protein phosphatases. To characterize its cellular role, PZL-1 was also expressed in Sz. pombe and in S. cerevisiae. Expression of PZL-1 in S. cerevisiae from the PPZ1 promoter was able to rescue the altered sensitivity to caffeine and lithium ions of a ppz1 strain. Furthermore, high copy number expression of PZL-1 alleviated the lytic phenotype of a S. cerevisiae slt2/mpk1 mitogen-activated protein (MAP) kinase mutant, similarly to that described for PPZ1, and mimicked the effects of high levels of Ppz1 on cell growth. Expression of PZL-1 in fission yeast from a weak version of the nmt1 promoter fully rescued the growth defect of a pzh1Delta strain in high potassium, but only partially complemented the sodium-hypertolerant phenotype. Strong overexpression of the N. crassa phosphatase in Sz. pombe affected cell growth and morphology. Therefore, PZL-1 appears to fulfil every known function carried out by its S. cerevisiae counterpart, despite the marked divergence in sequence within their NH(2)-terminal moieties.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalytic Domain
  • Cell Cycle
  • Cell Cycle Proteins*
  • Cloning, Molecular
  • Escherichia coli
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Gene Expression
  • Neurospora crassa / enzymology*
  • Neurospora crassa / genetics
  • Phosphoprotein Phosphatases / genetics*
  • Phosphoprotein Phosphatases / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins*
  • Schizosaccharomyces / genetics*

Substances

  • Cell Cycle Proteins
  • Fungal Proteins
  • SIS2 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • PPZ1 protein, S cerevisiae
  • Phosphoprotein Phosphatases