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Coordinate expression of NADPH-dependent flavin reductase, Fre-1, and Hint-related 7meGMP-directed hydrolase, DCS-1
Journal article   Open access   Peer reviewed

Coordinate expression of NADPH-dependent flavin reductase, Fre-1, and Hint-related 7meGMP-directed hydrolase, DCS-1

Dorota A Kwasnicka, Agnieszka Krakowiak, Colin Thacker, Charles Brenner and Steven R Vincent
The Journal of biological chemistry, Vol.278(40), pp.39051-39058
10/03/2003
DOI: 10.1074/jbc.M306355200
PMCID: PMC2556063
PMID: 12871939
url
https://doi.org/10.1074/jbc.M306355200View
Published (Version of record) Open Access

Abstract

A novel human cytosolic flavin reductase, Nr1, was recently described that contains FMN, FAD, and NADPH cofactors. Though the targets of the related NADPH-dependent flavoprotein reductases, cytochrome P450 reductase, methionine synthase reductase, and nitric oxide synthase, are known, the cellular function of Nr1 is not clear. To explore expression and regulation of Nr1, we cloned fre-1, the Caenorhabditis elegans ortholog of Nr1, and discovered that it is transcribed as a bicistronic pre-mRNA together with dcs-1, the ortholog of the recently described scavenger mRNA decapping enzyme. We used the novel substrate, 7meGpppBODIPY, to demonstrate that DCS-1 has low micromolar specificity for guanine ribonucleotides with the 7me modification, whereas trimethylated G substrates are poor competitors. Contrary to earlier classification, DCS-1 is not a pyrophosphatase but a distant member of the Hint branch of the histidine triad superfamily of nucleotide hydrolases and transferases. These observations are consistent with the hypothesis that DCS-1 homologs may function in the metabolism of capped oligonucleotides generated following exosome-dependent degradation of short-lived mRNA transcripts. We find that fre-1 and dcs-1 are coordinately expressed through worm development, are induced by heat shock, and have a nearly identical expression profile in human tissues. Furthermore, immunocytochemical analysis of the endogenous proteins in COS cells indicates that both are present in the nucleus and concentrated in a distinct perinuclear structure. Though no connection between these enzymes had been anticipated, our data and data from global expression and protein association studies suggest that the two enzymes jointly participate in responses to DNA damage, heat shock, and other stresses.
Immunohistochemistry Caenorhabditis elegans - chemistry Hydrolases - genetics Humans Molecular Sequence Data Saccharomyces cerevisiae Proteins - biosynthesis RNA, Messenger - metabolism N-Glycosyl Hydrolases - chemistry Tissue Distribution Cell Nucleus - metabolism Time Factors Cloning, Molecular N-Glycosyl Hydrolases - biosynthesis NADP - metabolism Recombinant Proteins - metabolism Amino Acid Sequence Boron Compounds - pharmacology Caenorhabditis elegans Proteins FMN Reductase - genetics Animals, Genetically Modified Operon Hot Temperature Reverse Transcriptase Polymerase Chain Reaction Pyrophosphatases - metabolism Sequence Homology, Amino Acid Animals FMN Reductase - chemistry Hydrolases - chemistry DNA Damage Kinetics Histidine - chemistry COS Cells Saccharomyces cerevisiae Proteins - chemistry

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