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Related Experiment Videos

Evidence for multiple nitrate uptake systems in the yeast Hansenula polymorpha.

F Machín1, G Perdomo, M D Pérez

  • 1Departamento de Bioquímica y Biología Molecular, Grupo del Metabolismo del Nitrógeno-Consejo Superior de Investigaciones Científicas, Universidad de La Laguna, E-38206 La Laguna, Tenerife, Canary Islands, Spain.

FEMS Microbiology Letters
|February 13, 2001
PubMed
Summary

Even without the main nitrate transporter (YNT1), Hansenula polymorpha can still absorb nitrate. A new system identified an additional nitrate uptake pathway, potentially also transporting nitrite.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Hansenula polymorpha utilizes nitrate as a nitrogen source, relying on specific transporter proteins.
  • The high-affinity nitrate transporter gene, YNT1, has been identified, but its absence doesn't fully abolish nitrate uptake.

Purpose of the Study:

  • To identify additional nitrate transporters in H. polymorpha responsible for nitrate uptake in YNT1 mutants.
  • To characterize the properties of this novel nitrate transport system.

Main Methods:

  • Construction of a modified H. polymorpha strain (FM31) with a disrupted nitrate assimilation gene cluster and a nitrate reductase gene (YNR1) under a strong promoter.
  • Quantification of nitrate uptake in the FM31 strain under various conditions.
  • Assessment of the influence of nitrogen sources (ammonium, glutamine) and inhibitors (chlorate) on nitrate uptake.

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Main Results:

  • Nitrate uptake was detected in the FM31 strain, independent of YNT1 and another transporter (Yna1p).
  • This uptake was not induced by nitrate or repressed by reduced nitrogen sources.
  • Inhibition of nitrite extrusion by external nitrite suggested the transporter might also mediate nitrite uptake.

Conclusions:

  • H. polymorpha possesses at least one additional nitrate uptake system beyond YNT1.
  • This newly identified system exhibits distinct regulation compared to known transporters.
  • The transporter may play a role in both nitrate and nitrite uptake, suggesting a broader role in nitrogen assimilation.