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: Keisuke Hikosaka, Masashi Ikutani, Masayuki Shito, Kohei Kazuma, Maryam Gulshan, Yoshinori Nagai, Kiyoshi Takatsu, Katsuhiro Konno, Kazuyuki Tobe, Hitoshi Kanno, Takashi Nakagawa
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 289 Issue 21 Pg. 14796-811 (May 23 2014) ISSN: 1083-351X [Electronic] United States
• PMID: 24739386 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• NAD
• Adenosine Triphosphate
• Nicotinamide-Nucleotide Adenylyltransferase
• nicotinamide mononucleotide adenylyltransferase 3, mouse

Topics

• Adenosine Triphosphate (metabolism)
• Anemia, Hemolytic (genetics, metabolism)
• Animals
• Blotting, Western
• Chromatography, Liquid
• Cytoplasm (enzymology)
• Erythrocytes (metabolism, ultrastructure)
• Glycolysis
• Metabolic Networks and Pathways (genetics)
• Metabolomics (methods)
• Mice
• Mice, Knockout
• Microscopy, Electron, Scanning
• NAD (metabolism)
• Nicotinamide-Nucleotide Adenylyltransferase (deficiency, genetics, metabolism)
• Splenomegaly (genetics, metabolism)
• Survival Analysis
• Tandem Mass Spectrometry