The terminal region of beta-catenin promotes stability by shielding the Armadillo repeats from the axin-scaffold destruction complex.
Post-translational stabilization of beta-catenin is a key step in Wnt signaling, but the features of beta-catenin required for stabilization are incompletely understood. We show that forms of beta-catenin lacking the unstructured C-terminal domain (CTD) show faster turnover than full-length or minimally truncated beta-catenins. Mutants that exhibit faster turnover show enhanced association with axin in co-transfected cells, and excess CTD polypeptide can compete binding of the beta-catenin armadillo (arm) repeat domain to axin in vitro, indicating that the CTD may restrict beta-catenin binding to the axin-scaffold complex. Fluorescent resonance energy transmission (FRET) analysis of cyan fluorescent protein (CFP)-arm-CTD-yellow fluorescent protein beta-catenin reveals that the CTD of beta-catenin can become spatially close to the N-terminal arm repeat region of beta-catenin. FRET activity is strongly diminished by the coexpression of beta-catenin binding partners, indicating that an unliganded groove is absolutely required for an orientation that allows FRET. Amino acids 733-759 are critical for beta-catenin FRET activity and stability. These data indicate that an N-terminal orientation of the CTD is required for beta-catenin stabilization and suggest a model where the CTD extends toward the N-terminal arm repeats, shielding these repeats from the beta-catenin destruction complex.
Mo
Rigen
R
Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611.
Chew
Teng-Leong
TL
Cellular and Molecular Biology, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611.
Maher
Meghan T
MT
Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611; Integrated Graduate Program in the Life Sciences, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611.
Bellipanni
Gianfranco
G
Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19106.
Weinberg
Eric S
ES
Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19106.
Gottardi
Cara J
CJ
Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611; Robert Lurie Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611. Electronic address: c-gottardi@northwestern.edu.
D000595:Amino Acid Sequence; D000818:Animals; D060466:Axin Protein; D002460:Cell Line; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D020798:Two-Hybrid System Techniques; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051176:beta Catenin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D060466:Axin Protein; D012097:Repressor Proteins; D051153:Wnt Proteins; D029961:Zebrafish Proteins; D051176:beta Catenin
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