Urmylation and tRNA thiolation functions of ubiquitin-like Uba4·Urm1 systems are conserved from yeast to man.
The ubiquitin-like protein Urm1 from budding yeast and its E1-like activator Uba4 have dual roles in protein urmylation and tRNA thiolation pathways. To study whether these are conserved among eukaryotes, we used gene shuffles to replace the yeast proteins by their human counterparts, hURM1 and hUBA4/MOCS3. As judged from biochemical and genetical assays, hURM1 and hUBA4 are functional in yeast, albeit at reduced efficiencies. They mediate urmylation of the peroxiredoxin Ahp1, a known urmylation target in yeast, and support tRNA thiolation. Similar to hUBA4, yeast Uba4 itself is modified by Urm1 and hURM1 suggesting target overlap between eukaryal urmylation pathways. In sum, our study shows that dual-function ubiquitin-like Urm1·Uba4 systems are conserved and exchangeable between human and yeast cells.
Universität Kassel, Institut für Biologie, FG Mikrobiologie, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.
André
A
Jüdes
Universität Kassel, Institut für Biologie, FG Mikrobiologie, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.
Folke
F
Ebert
Universität Kassel, Institut für Biologie, FG Mikrobiologie, Heinrich-Plett-Str. 40, 34132 Kassel, Germany; Spanish National Cancer Centre, Melchor Fernandez Almagro 3, Madrid, Spain.
Christian
C
Bär
Johannes Gutenberg Universität Mainz, Institut für Pharmazie und Biochemie, Staudinger Weg 5, 55128 Mainz, Germany.
Kathrin L
KL
Thüring
Universität Kassel, Institut für Biologie, FG Mikrobiologie, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.
Aileen
A
Harrer
Universität Kassel, Institut für Biologie, FG Mikrobiologie, Heinrich-Plett-Str. 40, 34132 Kassel, Germany.
Roland
R
Klassen
Johannes Gutenberg Universität Mainz, Institut für Pharmazie und Biochemie, Staudinger Weg 5, 55128 Mainz, Germany.
Mark
M
Helm
Centre for Gene Regulation & Expression, College of Life Sciences, MSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland, UK.
Michael J R
MJ
Stark
Universität Kassel, Institut für Biologie, FG Mikrobiologie, Heinrich-Plett-Str. 40, 34132 Kassel, Germany. Electronic address: schaffrath@uni-kassel.de.
Raffael
R
Schaffrath
D000926:Anticodon / Q000378:metabolism; D017124:Conserved Sequence; D006367:HeLa Cells; D006801:Humans; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000378:metabolism*; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013466:Sulfurtransferases / Q000737:chemistry / Q000378:metabolism*; D014452:Ubiquitins / Q000737:chemistry / Q000378:metabolism*
D000926:Anticodon; D029701:Saccharomyces cerevisiae Proteins; C406283:UBA4 protein, S cerevisiae; C406282:URM1 protein, S cerevisiae; D014452:Ubiquitins; C529881:Urm1 protein, human; D012343:RNA, Transfer; C502790:MOCS3 protein, human; D009713:Nucleotidyltransferases; D013466:Sulfurtransferases
Saccharomyces cerevisiae; Uba4 (hUBA4/MOCS3); Ubiquitin-like urmylation; Urm1 (hURM1); tRNA thiolation