The Structure of an LIM-Only Protein 4 (LMO4) and Deformed Epidermal Autoregulatory Factor-1 (DEAF1) Complex Reveals a Common Mode of Binding to LMO4

Abstract

LIM-domain only protein 4 (LMO4) is a widely expressed protein with important roles in embryonic development and breast cancer. It has been reported to bind many partners, including the transcription factor Deformed epidermal autoregulatory factor-1 (DEAF1), with which LMO4 shares many biological parallels. We used yeast two-hybrid assays to show that DEAF1 binds both LIM domains of LMO4 and that DEAF1 binds the same face on LMO4 as two other LMO4-binding partners, namely LIM domain binding protein 1 (LDB1) and C-terminal binding protein interacting protein (CtIP/RBBP8). Mutagenic screening analysed by the same method, indicates that the key residues in the interaction lie in LMO4LIM2 and the N-terminal half of the LMO4-binding domain in DEAF1. We generated a stable LMO4LIM2-DEAF1 complex and determined the solution structure of that complex. Although the LMO4-binding domain from DEAF1 is intrinsically disordered, it becomes structured on binding. The structure confirms that LDB1, CtIP and DEAF1 all bind to the same face on LMO4. LMO4 appears to form a hub in protein-protein interaction networks, linking numerous pathways within cells. Competitive binding for LMO4 therefore most likely provides a level of regulation between those different pathways.

hSSB1 (NABP2/OBFC2B) is regulated by oxidative stress

“…Size exclusion chromatography coupled to multi-angle laser light scattering was carried out as described previously 40 . Briefly, 250 μL (corresponding to 250 μg) of purified protein in MALS buffer (20 mM Tris pH 7, 100 mM NaCl, 1 mM EDTA, with or without 1 mM TCEP) was injected onto a Superose 12 10/30 analytical size exclusion column mounted to an AKTA chromatography system in tandem with a MALS detector.…”

Evidence for a Potential Common Gene Network of Smith-Magenis and Potocki-Lupski Syndromes, DAND (DEAF1-Associated Neurodevelopmental Disorder) and ZEB1-Associated Neurodevelopment Disorder

“…which also interact with the PI3K/mTOR pathway [55]. Yeast two-hybrid assays demonstrated that DEAF1 and LDB1 binds with LMO4 through LIM domains [56]. LDB1 interacts with CHD8, which is regulated by CTNNB1, a key downstream gene of GSK3B.…”

Preventing Jacob-induced transcriptional inactivation of CREB protects synapses from β-amyloid in Alzheimer’s Disease

“…2J). Previous studies suggest that binding regions for LMO4 frequently contain hydrophobic amino acids and particularly leucine (L) and valine (V) are crucial for the binding interface (34). Jacob contains a leucine-rich stretch of amino acids between 173-228 aa (Fig.…”