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The Ufm1-activating enzyme Uba5 is indispensable for erythroid differentiation in mice.

Tatsumi K, Yamamoto-Mukai H, Shimizu R, Waguri S, Sou YS, Sakamoto A, Taya C, Shitara H, Hara T, Chung CH, Tanaka K, Yamamoto M, Komatsu M - Nat Commun (2011)

Bottom Line: In this study, we report the essential role of Uba5, a specific activating enzyme for the ubiquitin-like modifier, Ufm1, in erythroid development.Although Uba5 was dispensable for the production of erythropoietin, its genetic loss led to impaired development of megakaryocyte and erythroid progenitors from common myeloid progenitors.Our results suggest that one of the ubiquitin-like protein modification systems, the Ufm1 system, is involved in the regulation of haematopoiesis.

View Article: PubMed Central - PubMed

Affiliation: Protein Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa 2-1-6, Setagaya-ku, Tokyo 156-8506, Japan.

ABSTRACT
Post-translational protein modifications are systems designed to expand restricted genomic information through functional conversion of target molecules. Ubiquitin-like post-translational modifiers regulate numerous cellular events through their covalent linkages to target protein(s) by an enzymatic cascade analogous to ubiquitylation consisting of E1 (activating), E2 (conjugating) and E3 (ligating) enzymes. In this study, we report the essential role of Uba5, a specific activating enzyme for the ubiquitin-like modifier, Ufm1, in erythroid development. Mice lacking Uba5 exhibited severe anaemia, followed by death in utero. Although Uba5 was dispensable for the production of erythropoietin, its genetic loss led to impaired development of megakaryocyte and erythroid progenitors from common myeloid progenitors. Intriguingly, transgenic expression of Uba5 in the erythroid lineage rescued the Uba5-deficient embryos from anaemia and prolonged their survival, demonstrating the importance of Uba5 in cell-autonomous erythroid differentiation. Our results suggest that one of the ubiquitin-like protein modification systems, the Ufm1 system, is involved in the regulation of haematopoiesis.

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Related in: MedlinePlus

Loss of Uba5 leads to death in utero because of severe anaemia.(a) Morphological examination of Uba5−/− mice embryos. Representative photographs of Uba5+/+ and Uba5−/− embryos at various developmental stages. Uba5−/− embryo at E12.5 exhibited severe anaemia. Scale bar, 2 mm. (b) Haematoxylin and eosin staining of fetal liver sections of Uba5+/+ and Uba5−/− mice at E11.5. Scale bars: left panel, 1 mm; right panel, 20 μm. (c) Number of erythrocytes in fetal livers of Uba5+/+ and Uba5−/− mice at E11.5. (d) Immunohistochemistry of fetal liver sections of Uba5+/+ and Uba5−/− embryos at E11.5 with anti-ɛ-globin and anti-β-globin antibodies. Arrows: abnormal multinucleated erythrocytes. Scale bar, 20 μm. (e) Representative photographs of Uba5+/+ and Uba5−/− embryos at E11.5. The circulating erythrocyte volume is reduced in the yolk sac of Uba5−/− embryos. Scale bar, 2 mm. (f) Wright–Giemsa staining of peripheral blood cytospin cells prepared from the indicated genotypes at E11.5. Scale bar, 20 μm. (g) Percentage of abnormal erythrocytes shown in f. Bar graphs in c and g show the mean±s.d. values of five mice from each group. Statistical analysis was carried out using the unpaired t-test. *P<0.05 (Welch test).
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f1: Loss of Uba5 leads to death in utero because of severe anaemia.(a) Morphological examination of Uba5−/− mice embryos. Representative photographs of Uba5+/+ and Uba5−/− embryos at various developmental stages. Uba5−/− embryo at E12.5 exhibited severe anaemia. Scale bar, 2 mm. (b) Haematoxylin and eosin staining of fetal liver sections of Uba5+/+ and Uba5−/− mice at E11.5. Scale bars: left panel, 1 mm; right panel, 20 μm. (c) Number of erythrocytes in fetal livers of Uba5+/+ and Uba5−/− mice at E11.5. (d) Immunohistochemistry of fetal liver sections of Uba5+/+ and Uba5−/− embryos at E11.5 with anti-ɛ-globin and anti-β-globin antibodies. Arrows: abnormal multinucleated erythrocytes. Scale bar, 20 μm. (e) Representative photographs of Uba5+/+ and Uba5−/− embryos at E11.5. The circulating erythrocyte volume is reduced in the yolk sac of Uba5−/− embryos. Scale bar, 2 mm. (f) Wright–Giemsa staining of peripheral blood cytospin cells prepared from the indicated genotypes at E11.5. Scale bar, 20 μm. (g) Percentage of abnormal erythrocytes shown in f. Bar graphs in c and g show the mean±s.d. values of five mice from each group. Statistical analysis was carried out using the unpaired t-test. *P<0.05 (Welch test).

Mentions: All Uba5−/− mice from Uba5+/− intercrosses died in utero, whereas Uba5-heterozygous (Uba5+/−) mice were born healthy and fertile without any noticeable pathological phenotype for at least 2 years. These results indicate that the complete loss of Uba5 is embryonically lethal. Analysis of Uba5−/− embryos at different developmental stages showed that the majority of embryos died between 12.5 and 13.5 embryonic days (E) after gestation (Table 1). The Uba5−/− embryos appeared paler and smaller than their wild-type littermates (Fig. 1a). The most obvious phenotype was severe fetal anaemia.


The Ufm1-activating enzyme Uba5 is indispensable for erythroid differentiation in mice.

Tatsumi K, Yamamoto-Mukai H, Shimizu R, Waguri S, Sou YS, Sakamoto A, Taya C, Shitara H, Hara T, Chung CH, Tanaka K, Yamamoto M, Komatsu M - Nat Commun (2011)

Loss of Uba5 leads to death in utero because of severe anaemia.(a) Morphological examination of Uba5−/− mice embryos. Representative photographs of Uba5+/+ and Uba5−/− embryos at various developmental stages. Uba5−/− embryo at E12.5 exhibited severe anaemia. Scale bar, 2 mm. (b) Haematoxylin and eosin staining of fetal liver sections of Uba5+/+ and Uba5−/− mice at E11.5. Scale bars: left panel, 1 mm; right panel, 20 μm. (c) Number of erythrocytes in fetal livers of Uba5+/+ and Uba5−/− mice at E11.5. (d) Immunohistochemistry of fetal liver sections of Uba5+/+ and Uba5−/− embryos at E11.5 with anti-ɛ-globin and anti-β-globin antibodies. Arrows: abnormal multinucleated erythrocytes. Scale bar, 20 μm. (e) Representative photographs of Uba5+/+ and Uba5−/− embryos at E11.5. The circulating erythrocyte volume is reduced in the yolk sac of Uba5−/− embryos. Scale bar, 2 mm. (f) Wright–Giemsa staining of peripheral blood cytospin cells prepared from the indicated genotypes at E11.5. Scale bar, 20 μm. (g) Percentage of abnormal erythrocytes shown in f. Bar graphs in c and g show the mean±s.d. values of five mice from each group. Statistical analysis was carried out using the unpaired t-test. *P<0.05 (Welch test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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f1: Loss of Uba5 leads to death in utero because of severe anaemia.(a) Morphological examination of Uba5−/− mice embryos. Representative photographs of Uba5+/+ and Uba5−/− embryos at various developmental stages. Uba5−/− embryo at E12.5 exhibited severe anaemia. Scale bar, 2 mm. (b) Haematoxylin and eosin staining of fetal liver sections of Uba5+/+ and Uba5−/− mice at E11.5. Scale bars: left panel, 1 mm; right panel, 20 μm. (c) Number of erythrocytes in fetal livers of Uba5+/+ and Uba5−/− mice at E11.5. (d) Immunohistochemistry of fetal liver sections of Uba5+/+ and Uba5−/− embryos at E11.5 with anti-ɛ-globin and anti-β-globin antibodies. Arrows: abnormal multinucleated erythrocytes. Scale bar, 20 μm. (e) Representative photographs of Uba5+/+ and Uba5−/− embryos at E11.5. The circulating erythrocyte volume is reduced in the yolk sac of Uba5−/− embryos. Scale bar, 2 mm. (f) Wright–Giemsa staining of peripheral blood cytospin cells prepared from the indicated genotypes at E11.5. Scale bar, 20 μm. (g) Percentage of abnormal erythrocytes shown in f. Bar graphs in c and g show the mean±s.d. values of five mice from each group. Statistical analysis was carried out using the unpaired t-test. *P<0.05 (Welch test).
Mentions: All Uba5−/− mice from Uba5+/− intercrosses died in utero, whereas Uba5-heterozygous (Uba5+/−) mice were born healthy and fertile without any noticeable pathological phenotype for at least 2 years. These results indicate that the complete loss of Uba5 is embryonically lethal. Analysis of Uba5−/− embryos at different developmental stages showed that the majority of embryos died between 12.5 and 13.5 embryonic days (E) after gestation (Table 1). The Uba5−/− embryos appeared paler and smaller than their wild-type littermates (Fig. 1a). The most obvious phenotype was severe fetal anaemia.

Bottom Line: In this study, we report the essential role of Uba5, a specific activating enzyme for the ubiquitin-like modifier, Ufm1, in erythroid development.Although Uba5 was dispensable for the production of erythropoietin, its genetic loss led to impaired development of megakaryocyte and erythroid progenitors from common myeloid progenitors.Our results suggest that one of the ubiquitin-like protein modification systems, the Ufm1 system, is involved in the regulation of haematopoiesis.

View Article: PubMed Central - PubMed

Affiliation: Protein Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Kamikitazawa 2-1-6, Setagaya-ku, Tokyo 156-8506, Japan.

ABSTRACT
Post-translational protein modifications are systems designed to expand restricted genomic information through functional conversion of target molecules. Ubiquitin-like post-translational modifiers regulate numerous cellular events through their covalent linkages to target protein(s) by an enzymatic cascade analogous to ubiquitylation consisting of E1 (activating), E2 (conjugating) and E3 (ligating) enzymes. In this study, we report the essential role of Uba5, a specific activating enzyme for the ubiquitin-like modifier, Ufm1, in erythroid development. Mice lacking Uba5 exhibited severe anaemia, followed by death in utero. Although Uba5 was dispensable for the production of erythropoietin, its genetic loss led to impaired development of megakaryocyte and erythroid progenitors from common myeloid progenitors. Intriguingly, transgenic expression of Uba5 in the erythroid lineage rescued the Uba5-deficient embryos from anaemia and prolonged their survival, demonstrating the importance of Uba5 in cell-autonomous erythroid differentiation. Our results suggest that one of the ubiquitin-like protein modification systems, the Ufm1 system, is involved in the regulation of haematopoiesis.

Show MeSH
Related in: MedlinePlus