Hub1 is an essential ubiquitin-like protein without functioning as a typical modifier in fission yeast.
Hub1 exhibits 23% sequence identity to ubiquitin. However, Hub1 lacks the C-terminal Gly, which is essential for covalent attachment to target protein(s) of ubiquitin and other ubiquitin-like (UBL) modifiers. Instead, Hub1 proteins in all eukaryotes retain the di-Tyr just before a single variable residue at the C-terminus, so one intriguing question is whether Hub1 could be linked to substrate through the conserved Tyr or not. Here we studied Hub1 in Schizosaccharomyces pombe. Gene disruption experiment revealed that hub1+ is essential. Remarkably, the mutant cells harbouring Hub1 lacking the di-Tyr could grow similar to wild-type cells, indicating that the di-Tyr is dispensable for the essential function of Hub1. Moreover, we could not observe cleavage of Flag-tag fused with C-terminus of Hub1. It suggests that the processing for conjugation via conserved Tyr is not likely to occur in Hub1, and Hub1 is a novel class of the UBL protein family. Finally, we isolated a temperature-sensitive allele, hub1-1. This temperature sensitivity could be suppressed by overproduction of Rpb10 or Snu66, the former of which is one of the common subunits of the RNA polymerases and the other is the component of the spliceosome. We also observed that pre-mRNA splicing was impaired in hub1-1.
Yashiroda
Hideki
H
Department of Molecular Oncology, Tokyo Metropolitan Institute of Medical Science, Honkomagome 3-18-22, Bunkyo-ku, Tokyo 113-8613, Japan. hyashiro@rinshoken.or.jp
Tanaka
Keiji
K
D000483:Alleles; D000595:Amino Acid Sequence; D006801:Humans; D009154:Mutation; D046188:Protein Modification, Translational; D012568:Schizosaccharomyces / Q000737:chemistry / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D016415:Sequence Alignment; D013696:Temperature; D043743:Ubiquitin-Protein Ligase Complexes / Q000235:genetics / Q000502:physiology*; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism*
C500017:HUB1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D014452:Ubiquitins; D043743:Ubiquitin-Protein Ligase Complexes