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

Emese Vissi, Josep Clotet, Eulalia de Nadal, Anna Barceló, Éva Bakó, Pál Gergely, Viktor Dombrádi, Joaquín Ariño

Research output: Contribution to journalArticle

19 Citations (Scopus)

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 pzh1Δ 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 NH2-terminal moieties.

Original languageEnglish
Pages (from-to)115-124
Number of pages10
JournalYeast
Volume18
Issue number2
DOIs
Publication statusPublished - Jan 30 2001

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Keywords

  • Functional expression
  • Ppz phosphatases
  • Salt tolerance
  • Yeast

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Genetics

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