Mutations in Protein-Binding Hot-Spots on the Hub Protein Smad3 Differentially Affect Its Protein Interactions and Smad3-Regulated Gene Expression

Hub proteins are connected through binding interactions to many other proteins. Smad3, a mediator of signal transduction induced by transforming growth factor beta (TGF-β), serves as a hub protein for over 50 protein-protein interactions. Different cellular responses mediated by Smad3 are the product of cell-type and context dependent Smad3-nucleated protein complexes acting in concert. Our hypothesis is that perturbation of this spectrum of protein complexes by mutation of single protein-binding hot-spots on Smad3 will have distinct consequences on Smad3-mediated responses.We mutated 28 amino acids on the surface of the Smad3 MH2 domain and identified 22 Smad3 variants with reduced binding to subsets of 17 Smad3-binding proteins including Smad4, SARA, Ski, Smurf2 and SIP1. Mutations defective in binding to Smad4, e.g., D408H, or defective in nucleocytoplasmic shuttling, e.g., W406A, were compromised in modulating the expression levels of a Smad3-dependent reporter gene or six endogenous Smad3-responsive genes: Mmp9, IL11, Tnfaip6, Fermt1, Olfm2 and Wnt11. However, the Smad3 mutants Y226A, Y297A, W326A, K341A, and E267A had distinct differences on TGF-β signaling. For example, K341A and Y226A both reduced the Smad3-mediated activation of the reporter gene by ∼50% but K341A only reduced the TGF-β inducibilty of Olfm2 in contrast to Y226A which reduced the TGF-β inducibility of all six endogenous genes as severely as the W406A mutation. E267A had increased protein binding but reduced TGF-β inducibility because it caused higher basal levels of expression. Y297A had increased TGF-β inducibility because it caused lower Smad3-induced basal levels of gene expression.Mutations in protein binding hot-spots on Smad3 reduced the binding to different subsets of interacting proteins and caused a range of quantitative changes in the expression of genes induced by Smad3. This approach should be useful for unraveling which Smad3 protein complexes are critical for specific biological responses

Accumulators in (and Beyond) Generic Groups: Non-Trivial Batch Verification Requires Interaction

We prove a tight lower bound on the number of group operations required for batch verification by any generic-group accumulator that stores a less-than-trivial amount of information. Specifically, we show that $\Omega(t \cdot (\lambda / \log \lambda))$ group operations are required for the batch verification of any subset of $t \geq 1$ elements, where $\lambda \in \mathbb{N}$ is the security parameter, thus ruling out non-trivial batch verification in the standard non-interactive manner. Our lower bound applies already to the most basic form of accumulators (i.e., static accumulators that support membership proofs), and holds both for known-order (and even multilinear) groups and for unknown-order groups, where it matches the asymptotic performance of the known bilinear and RSA accumulators, respectively. In addition, it complements the techniques underlying the generic-group accumulators of Boneh, B{ü}nz and Fisch (CRYPTO \u2719) and Thakur (ePrint \u2719) by justifying their application of the Fiat-Shamir heuristic for transforming their interactive batch-verification protocols into non-interactive procedures. Moreover, motivated by a fundamental challenge introduced by Aggarwal and Maurer (EUROCRYPT \u2709), we propose an extension of the generic-group model that enables us to capture a bounded amount of arbitrary non-generic information (e.g., least-significant bits or Jacobi symbols that are hard to compute generically but are easy to compute non-generically). We prove our lower bound within this extended model, which may be of independent interest for strengthening the implications of impossibility results in idealized models

The Ski proto-oncogene regulates body composition and suppresses lipogenesis

Objective: The Ski gene regulates skeletal muscle differentiation in vitro and and in vivo. In the c-Ski overexpression mouse model there occurs marked skeletal muscle hypertrophy with decreased adipose tissue mass. In this study, we have investigated the underlying molecular mechanisms responsible for the increased skeletal muscle and decreased adipose tissue mass in the c-Ski mouse