Protoplasma

, 238:59 | Cite as

Nitric oxide as a signaling molecule in the fission yeast Schizosaccharomyces pombe

Original Article
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Abstract

Nitric oxide synthases (NOS) catalyze the synthesis of ubiquitous signaling molecule nitric oxide (NO) which controls numerous biological processes. Using a spectrofluorometric NOS assay, we have measured the rate of total NO production in the crude cell extracts of Schizosaccharomyces pombe. NO production was reduced in the absence of NOS cofactors calmodulin and tetrahydrobiopterin, and a competitive NOS inhibitor NG-nitro-l-arginine methyl ester (l-NAME) was able to cause a statistically significant inhibition on the rate of total NO production. These results, for the first time, provide evidence that an enzyme with a NOS-like activity may be present in the fission yeast. In order to assess the possible regulatory roles of NO as a signaling molecule in this yeast, using the differential display technique, we screened for NO-responsive genes whose expression decreased upon exposure to l-NAME and increased in response to an NO donor, sodium nitroprusside treatment. Differential expression patterns of byr1, pek1, sid1, and wis1 genes were confirmed by quantitative real-time PCR. The physiological experiments performed based on the functions and molecular interactions of these genes have pointed to the possibility that NO production might be required for sporulation in S. pombe. Taken together, these findings suggest that NO may function as a signaling molecule which can induce both transcriptional and physiological changes in the fission yeast. Hence, these data also imply that S. pombe can be used as a model system for investigating the mechanisms underlying NO-related complex signaling pathways.

Keywords

Schizosaccharomyces pombe Nitric oxide (NO) Differential display Nitric oxide synthase (NOS) Sporulation 

Abbreviations

ANOVA

Analysis of variance

BH4

Tetrahydrobiopterin

BLAST

Basic local alignment search tool

bp

Base pair

CaM

Calmodulin

DAF-2

4,5-Diaminofluorescein

DAF-2T

4,5-Diaminofluorescein triazole

d-NAME

NG-Nitro-d-arginine methyl ester

GC

Guanylyl cyclase

l-NAME

NG-Nitro-l-arginine methyl ester

MAPK

Mitogen-activated protein kinase

MEA

Malt extract agar

MMA

Minimal medium agar

MML

Minimal medium liquid

NO

Nitric oxide

NOS

Nitric oxide synthase

ODQ

1H-[1,2,4]Oxadiazole[4,3-a]quinoxalin-1-one

PAK

p21-activated kinase

SAPK

Stress-activated protein kinase

SNP

Sodium nitroprusside

YEA

Yeast extract agar

YEL

Yeast extract liquid

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Notes

Acknowledgments

This study was supported by the Research Fund of Istanbul University (project no. T-747/13092005).

Conflict of interest statement

I, Prof. Dr. Guler Temizkan, declare that I have no conflict of interest with the Research Fund of Istanbul University.

I, Dr. Cenk Kig, declare that I have no conflict of interest with the Research Fund of Istanbul University.

Supplementary material

709_2009_74_MOESM1_ESM.pdf (268 kb)
Supplementary data (PDF 274 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Molecular Biology and Genetics, Faculty of ScienceIstanbul UniversityIstanbulTurkey

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