E6AP ubiquitin ligase mediates ubiquitin-dependent degradation of peroxiredoxin 1.
E6-associated protein (E6AP) is a cellular ubiquitin protein ligase that mediates ubiquitylation and degradation of tumor suppressor p53 in conjunction with the high-risk human papillomavirus E6 protein. We previously reported that E6AP targets annexin A1 protein for ubiquitin-dependent proteasomal degradation. To gain a better understanding of the physiological function of E6AP, we have been seeking to identify novel substrates of E6AP. Here, we identified peroxiredoxin 1 (Prx1) as a novel E6AP-binding protein using a tandem affinity purification procedure coupled with mass spectrometry. Prx1 is a 25-kDa member of the Prx family, a ubiquitous family of antioxidant peroxidases that regulate many cellular processes through intracellular oxidative signal transduction pathways. Immunoprecipitation analysis showed that E6AP binds Prx1 in vivo. Pull-down experiments showed that E6AP binds Prx1 in vitro. Ectopic expression of E6AP enhanced the degradation of Prx1 in vivo. In vivo and in vitro ubiquitylation assays revealed that E6AP promoted polyubiquitylation of Prx1. RNAi-mediated downregulation of endogenous E6AP increased the level of endogenous Prx1 protein. Taken together, our data suggest that E6AP mediates the ubiquitin-dependent proteasomal degradation of Prx1. Our findings raise a possibility that E6AP may play a role in regulating Prx1-dependent intracellular oxidative signal transduction pathways.
Nasu
Junichi
J
Department of Virology II, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan.
Murakami
Kyoko
K
Miyagawa
Shoji
S
Yamashita
Ryosuke
R
Ichimura
Tohru
T
Wakita
Takaji
T
Hotta
Hak
H
Miyamura
Tatsuo
T
Suzuki
Tetsuro
T
Satoh
Tazuko
T
Shoji
Ikuo
I
D000975:Antioxidants; D002460:Cell Line; D006801:Humans; D010084:Oxidation-Reduction; D054464:Peroxiredoxins / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction; D025801:Ubiquitin; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
D000975:Antioxidants; D025801:Ubiquitin; D054464:Peroxiredoxins; C471639:UBE3A protein, human; D044767:Ubiquitin-Protein Ligases