The Pneumocystis meiotic PCRan1p kinase exhibits unique temperature-regulated activity

Joshua W. Burgess, Theodore J. Kottom, Leah R. Villegas, Jeffrey D. Lamont, Elizabeth M. Baden, Marina Ramirez-Alvarado, Andrew Harold Limper

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Pneumocystis organisms are opportunistic fungal pathogens that cause significant pneumonia in immune-compromised hosts. Recent evidence has suggested that Pneumocystis carinii exists as separate mating types, and expresses and regulates proteins that govern meiosis and progression of the life cycle. This study was undertaken to investigate the activity of three life cycle-regulatory proteins in Pneumocystis, including two proteins essential in mating signaling, and a putative meiotic regulator, to determine the conditions under which they are mostactive. This study used V5/HIS-tagged PCRan1p, PCSte20p, and PCCbk1, purified from Saccharomyces cerevisiae strain, INVSC, as well as an in vitro Escherichia coli protein expression system to determine the optimal expression conditions of each protein in the presence of varying pH, temperature, and metal ions. These studies demonstrate an atypical enzymatic activity in PCRan1p, whereby the kinase was most active in the environmental conditions between 10 and 25°C, compared with a dramatic reduction in activity above 30°C, temperatures typically found within mammalianhosts. Circular dichroism andfluorescence spectroscopy suggest that PCRan1p becomespartially unfoldedat 25°C, leading to its most active conformation, whereas continued unfolding as temperature increases results in strongly suppressed activity. These studies suggest that, in vivo, while under conditions within the mammalian lung (typically 37°C), PCRan1p kinase activity is largely suppressed, allowing better conditions for the activation of meiosis, whereas in ex vivo environments, PCRan1p kinase activity increases to arrest progression of the life cycle until conditions become more favorable.

Original languageEnglish (US)
Pages (from-to)714-721
Number of pages8
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume41
Issue number6
DOIs
StatePublished - Dec 1 2009

Fingerprint

Pneumocystis
Phosphotransferases
Life Cycle Stages
Life cycle
Temperature
Meiosis
Proteins
Circular dichroism spectroscopy
Pneumocystis carinii
Escherichia coli Proteins
Pathogens
Circular Dichroism
Yeast
Metal ions
Saccharomyces cerevisiae
Conformations
Spectrum Analysis
Pneumonia
Thermodynamic properties
Metals

Keywords

  • Kinase
  • Meiosis
  • Pneumocystis
  • Temperature

ASJC Scopus subject areas

  • Cell Biology
  • Pulmonary and Respiratory Medicine
  • Molecular Biology
  • Clinical Biochemistry

Cite this

The Pneumocystis meiotic PCRan1p kinase exhibits unique temperature-regulated activity. / Burgess, Joshua W.; Kottom, Theodore J.; Villegas, Leah R.; Lamont, Jeffrey D.; Baden, Elizabeth M.; Ramirez-Alvarado, Marina; Limper, Andrew Harold.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 41, No. 6, 01.12.2009, p. 714-721.

Research output: Contribution to journalArticle

Burgess, Joshua W. ; Kottom, Theodore J. ; Villegas, Leah R. ; Lamont, Jeffrey D. ; Baden, Elizabeth M. ; Ramirez-Alvarado, Marina ; Limper, Andrew Harold. / The Pneumocystis meiotic PCRan1p kinase exhibits unique temperature-regulated activity. In: American Journal of Respiratory Cell and Molecular Biology. 2009 ; Vol. 41, No. 6. pp. 714-721.
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