title: RCAD/Ufl1, a Ufm1 E3 ligase, is essential for hematopoietic stem cell function and murine hematopoiesis.
abstract: The Ufm1 conjugation system is a novel ubiquitin-like modification system, consisting of Ufm1, Uba5 (E1), Ufc1 (E2) and poorly characterized E3 ligase(s). RCAD/Ufl1 (also known as KIAA0776, NLBP and Maxer) was reported to function as a Ufm1 E3 ligase in ufmylation (Ufm1-mediated conjugation) of DDRGK1 and ASC1 proteins. It has also been implicated in estrogen receptor signaling, unfolded protein response (UPR) and neurodegeneration, yet its physiological function remains completely unknown. In this study, we report that RCAD/Ufl1 is essential for embryonic development, hematopoietic stem cell (HSC) survival and erythroid differentiation. Both germ-line and somatic deletion of RCAD/Ufl1 impaired hematopoietic development, resulting in severe anemia, cytopenia and ultimately animal death. Depletion of RCAD/Ufl1 caused elevated endoplasmic reticulum stress and evoked UPR in bone marrow cells. In addition, loss of RCAD/Ufl1 blocked autophagic degradation, increased mitochondrial mass and reactive oxygen species, and led to DNA damage response, p53 activation and enhanced cell death of HSCs. Collectively, our study provides the first genetic evidence for the indispensable role of RCAD/Ufl1 in murine hematopoiesis and development. The finding of RCAD/Ufl1 as a key regulator of cellular stress response sheds a light into the role of a novel protein network including RCAD/Ufl1 and its associated proteins in regulating cellular homeostasis.
authors.affiliation: Department of Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, China.|Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: M
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.|Department of Orthopedic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.|Department of Biology, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Sivaprakasam
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Gurav
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: W
authors.initials: W
authors.lastname: Pi
authors.affiliation: Department of Pediatrics, Georgia Regents University, Augusta, GA, USA.
authors.forename: L
authors.initials: L
authors.lastname: Makala
authors.affiliation: Department of Periodontics, College of Dentistry, Cancer Center, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: J
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.|Department of Pediatrics, Georgia Regents University, Augusta, GA, USA.
authors.forename: B
authors.initials: B
authors.lastname: Pace
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: D
authors.initials: D
authors.lastname: Tuan-Lo
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: V
authors.initials: V
authors.lastname: Ganapathy
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: N
authors.initials: N
authors.lastname: Singh
authors.affiliation: Department of Biochemistry & Molecular Biology, Cancer Center, Georgia Regents University, Augusta, GA, USA.
authors.forename: H
authors.initials: H
authors.lastname: Li
mesh_terms: D000740:Anemia / Q000209:etiology; D000818:Animals; D001343:Autophagy; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D059865:Endoplasmic Reticulum Stress; D057809:HEK293 Cells; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D034622:RNA Interference; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D056811:Unfolded Protein Response
chemical_list: D008869:Microtubule-Associated Proteins; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D044767:Ubiquitin-Protein Ligases