title: A novel approach to assess the ubiquitin-fold modifier 1-system in cells.
abstract: The ubiquitin-fold modifier 1 (UFM1)-system, a ubiquitin-like protein conjugation system, is involved in the development of breast cancer and several hereditary neurological syndromes. However, the molecular mechanisms of UFM1-related pathogenesis remain unclear. Here, we show that in the absence of UFSP2, a deconjugating enzyme for UFM1, ectopic expression of both UFL1 and UFBP1, which serve as the E3-ligase complex for the UFM1-system, dramatically increases UFM1-conjugate formation at the endoplasmic reticulum. Utilizing this system, we were able to attribute disease-related isoforms of UBA5, the E1 enzyme for UFM1, to decreased UFM1-conjugate formation. Our procedure allows the assessment of UFM1-conjugate formation in cells and the identification of UFM1-targets, both of which are needed to clarify the pathophysiological role of the UFM1-system.
authors.affiliation: Department of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences, Japan.|Laboratory of Protein Metabolism, The Tokyo Metropolitan Institute of Medical Science, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Ishimura
authors.affiliation: Department of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Obata
authors.affiliation: Department of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Shun
authors.initials: S
authors.lastname: Kageyama
authors.affiliation: Department of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Jens
authors.initials: J
authors.lastname: Daniel
authors.affiliation: Laboratory of Protein Metabolism, The Tokyo Metropolitan Institute of Medical Science, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: Department of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Komatsu
mesh_terms: D001681:Biological Assay / Q000379:methods*; D055786:Gene Knockout Techniques; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D011506:Proteins / Q000378:metabolism*; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism
chemical_list: D011506:Proteins; C502094:UFM1 protein, human; D014452:Ubiquitins
keywords: UFM1; encephalopathy; ubiquitin-like modifier