The Polycomb protein and E3 ubiquitin ligase Ring1B harbors an IRES in its highly conserved 5' UTR.
Ring1B is an essential member of the highly conserved Polycomb group proteins, which orchestrate developmental processes, cell growth and stem cell fate by modifying local chromatin structure. Ring1B was found to be the E3 ligase that monoubiquitinates histone H2A, which adds a new level of chromatin modification to Polycomb group proteins. Here we report that Ring1B belongs to the exclusive group of proteins that for their translation depend on a stable 5' UTR sequence in their mRNA known as an Internal Ribosome Entry Site (IRES). In cell transfection assays the Ring1B IRES confers significantly higher expression levels of Ring1B than a Ring1B cDNA without the IRES. Also, dual luciferase assays show strong activity of the Ring1B IRES. Although our findings indicate Ring1B can be translated under conditions where cap-dependent translation is impaired, we found the Ring1B IRES to be cap-dependent. This raises the possibility that translational control of Ring1B is a multi-layered process and that translation of Ring1B needs to be maintained under varying conditions, which is in line with its essential role as an E3 ligase for monoubiquitination of histone H2A in the PRC1 Polycomb protein complex.
Division of Molecular Genetics, Netherlands Cancer Institute, Amsterdam, The Netherlands.
Erwin
E
Boutsma
Sonja
S
Noback
Maarten
M
van Lohuizen
D020121:5' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D017931:DNA Primers; D020224:Expressed Sequence Tags; D017354:Point Mutation; D063150:Polycomb Repressive Complex 1; D063146:Polycomb-Group Proteins; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics; D012270:Ribosomes; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000235:genetics
D020121:5' Untranslated Regions; D017931:DNA Primers; D063146:Polycomb-Group Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D063150:Polycomb Repressive Complex 1; D044767:Ubiquitin-Protein Ligases