title: Deficiency of nicotinamide mononucleotide adenylyltransferase 3 (nmnat3) causes hemolytic anemia by altering the glycolytic flow in mature erythrocytes.
abstract: NAD biosynthesis is of substantial interest because of its important roles in regulating various biological processes. Nicotinamide mononucleotide adenylyltransferase 3 (Nmnat3) is considered a mitochondria-localized NAD synthesis enzyme involved in de novo and salvage pathways. Although the biochemical properties of Nmnat3 are well documented, its physiological function in vivo remains unclear. In this study, we demonstrated that Nmnat3 was localized in the cytoplasm of mature erythrocytes and critically regulated their NAD pool. Deficiency of Nmnat3 in mice caused splenomegaly and hemolytic anemia, which was associated with the findings that Nmnat3-deficient erythrocytes had markedly lower ATP levels and shortened lifespans. However, the NAD level in other tissues were not apparently affected by the deficiency of Nmnat3. LC-MS/MS-based metabolomics revealed that the glycolysis pathway in Nmnat3-deficient erythrocytes was blocked at a glyceraldehyde 3-phosphate dehydrogenase (GAPDH) step because of the shortage of the coenzyme NAD. Stable isotope tracer analysis further demonstrated that deficiency of Nmnat3 resulted in glycolysis stall and a shift to the pentose phosphate pathway. Our findings indicate the critical roles of Nmnat3 in maintenance of the NAD pool in mature erythrocytes and the physiological impacts at its absence in mice.
authors.affiliation: From the Frontier Research Core for Life Sciences.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Hikosaka
authors.affiliation: Department of Immunobiology and Pharmacological Genetics, Graduate School of Medicine and Pharmaceutical Science for Research.
authors.forename: Masashi
authors.initials: M
authors.lastname: Ikutani
authors.affiliation: the Departments of Transfusion Medicine and Cell Processing and.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Shito
authors.affiliation: the Institute of Natural Medicine, University of Toyama, Toyama 930-0194.
authors.forename: Kohei
authors.initials: K
authors.lastname: Kazuma
authors.affiliation: From the Frontier Research Core for Life Sciences, The First Department of Internal Medicine, Graduate School of Medicine and Pharmaceutical Science for Research, and.
authors.forename: Maryam
authors.initials: M
authors.lastname: Gulshan
authors.affiliation: Department of Immunobiology and Pharmacological Genetics, Graduate School of Medicine and Pharmaceutical Science for Research, the JST, PRESTO, Saitama 332-0012, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Nagai
authors.affiliation: Department of Immunobiology and Pharmacological Genetics, Graduate School of Medicine and Pharmaceutical Science for Research, the Toyama Prefectural Institute for Pharmaceutical Research, Toyama 939-0363, and.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Takatsu
authors.affiliation: the Institute of Natural Medicine, University of Toyama, Toyama 930-0194.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Konno
authors.affiliation: The First Department of Internal Medicine, Graduate School of Medicine and Pharmaceutical Science for Research, and.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Tobe
authors.affiliation: the Departments of Transfusion Medicine and Cell Processing and Advanced Biomedical Engineering and Science, Graduate School of Medicine, Tokyo Women's Medical University, Tokyo 162-8666.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kanno
authors.affiliation: From the Frontier Research Core for Life Sciences, nakagawa@med.u-toyama.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakagawa
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000743:Anemia, Hemolytic / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D002853:Chromatography, Liquid; D003593:Cytoplasm / Q000201:enzymology; D004912:Erythrocytes / Q000378:metabolism* / Q000648:ultrastructure; D006019:Glycolysis; D053858:Metabolic Networks and Pathways / Q000235:genetics; D055432:Metabolomics / Q000379:methods; D051379:Mice; D018345:Mice, Knockout; D008855:Microscopy, Electron, Scanning; D009243:NAD / Q000378:metabolism; D009612:Nicotinamide-Nucleotide Adenylyltransferase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D013163:Splenomegaly / Q000235:genetics / Q000378:metabolism; D016019:Survival Analysis; D053719:Tandem Mass Spectrometry
chemical_list: D009243:NAD; D000255:Adenosine Triphosphate; D009612:Nicotinamide-Nucleotide Adenylyltransferase; C000592579:nicotinamide mononucleotide adenylyltransferase 3, mouse
keywords: Erythrocyte; Glycolysis; Metabolomics; NAD Biosynthesis; Nicotinamide Adenine Dinucleotide (NAD)
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