title: Ubiquitination of an artificial RING finger without a substrate and a tag.
abstract: Alpha-helical region substitution was applied to the SIAH1 and EL5 RING fingers. The Williams-Beuren syndrome transcription factor (WSTF) PHD_SIAH1 and WSTF PHD_EL5 RING fingers were created as the artificial ubiquitin-ligating enzyme (E3). These fingers possess E3 activities of mono-ubiquitination and poly-ubiquitination, respectively, with ubiquitin-conjugating enzyme (E2)-binding capabilities. Artificial E3s bind two zinc atoms and adopt a zinc-dependent ordered structure and ubiquitinate upon themselves without a substrate and a tag. Ubiquitination experiments using biotinylated ubiquitin showed that the WSTF PHD_EL5 RING finger is poly-ubiquitinated via residue Lys(63) of ubiquitin. Substitution of alpha-helical region might be applicable to various RING fingers with mono-ubiquitination or poly-ubiquitination.
authors: Department of Pharmaceutical Health Care, Himeji Dokkyo University, Himeji, Hyogo, Japan. miyamoto@himeji-du.ac.jp
authors: Kazuhide
authors: K
authors: Miyamoto
mesh_terms: D000595:Amino Acid Sequence; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D025821:Polyubiquitin / Q000737:chemistry; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary; D054829:RING Finger Domains; D013379:Substrate Specificity; D014157:Transcription Factors / Q000138:chemical synthesis / Q000737:chemistry*; D044763:Ubiquitin-Conjugating Enzymes / Q000138:chemical synthesis / Q000737:chemistry; D044767:Ubiquitin-Protein Ligases / Q000138:chemical synthesis / Q000737:chemistry; D054875:Ubiquitination; D015032:Zinc / Q000737:chemistry; D000092468:Bromodomain Containing Proteins
chemical_list: D010455:Peptides; D025821:Polyubiquitin; D014157:Transcription Factors; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases; D015032:Zinc; C116750:BAZ1B protein, human; D000092468:Bromodomain Containing Proteins