ABSTRACT
The effects of cladding layers of rate-sensitive materials on the ductility and fracture strain of compressed rings are numerically investigated by using the finite element method (FEM) and employing the Johnson-Cook (J-C) model. The results show that ductility is governed by the behavior of the material that is located at the ring outer wall regardless of the volume fraction of the core and clad materials. However, as the number of layers increases, this influence becomes less noticeable. Moreover, as barreling increases at the outer wall and decreases at the inner wall, fracture strain increases. Furthermore, the effects of ring shape factor and bonding type of clad and core materials are numerically evaluated. The numerical results show that less force per unit volume is required to fracture narrower rings and that using a noise diffusion pattern at the interface of the materials is more suitable to simulate crack propagation in the compressed rings and functionally graded materials (FGMs). Additionally, delamination has a direct relation to layer thickness and can occur even in the presence of perfect bonding conditions owing to differences among the material and fracture parameters of laminated layers.
ABSTRACT
RasGRPs (guanine-nucleotide-releasing proteins) are exchange factors for membrane-bound GTPases. All RasGRP family members contain C1 domains which, in other proteins, bind DAG (diacylglycerol) and thus mediate the proximal signal-transduction events induced by this lipid second messenger. The presence of C1 domains suggests that all RasGRPs could be regulated by membrane translocation driven by C1-DAG interactions. This has been demonstrated for RasGRP1 and RasGRP3, but has not been tested directly for RasGRP2, RasGRP4alpha and RasGRP4beta. Sequence alignments indicate that all RasGRP C1 domains have the potential to bind DAG. In cells, the isolated C1 domains of RasGRP1, RasGRP3 and RasGRP4alpha co-localize with membranes and relocalize in response to DAG, whereas the C1 domains of RasGRP2 and RasGRP4beta do not. Only the C1 domains of RasGRP1, RasGRP3 and RasGRP4alpha recognize DAG as a ligand within phospholipid vesicles and do so with differential affinities. Other lipid second messengers were screened as ligands for RasGRP C1 domains, but none was found to serve as an alternative to DAG. All of the RasGRP C1 domains bound to vesicles which contained a high concentration of anionic phospholipids, indicating that this could provide a DAG-independent mechanism for membrane binding by C1 domains. This concept was supported by demonstrating that the C1 domain of RasGRP2 could functionally replace the membrane-binding role of the C1 domain within RasGRP1, despite the inability of the RasGRP2 C1 domain to bind DAG. The RasGRP4beta C1 domain was non-functional when inserted into either RasGRP1 or RasGRP4, implying that the alternative splicing which produces this C1 domain eliminates its contribution to membrane binding.
Subject(s)
Cell Membrane/metabolism , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/metabolism , Diglycerides/chemistry , Diglycerides/metabolism , Guanine Nucleotide Exchange Factors/chemistry , Guanine Nucleotide Exchange Factors/metabolism , Amino Acid Sequence , Animals , Anions/chemistry , Cell Line , Humans , Mice , Models, Molecular , Molecular Sequence Data , Mutation/genetics , Phorbol Esters , Phospholipids/chemistry , Protein Structure, Tertiary , Protein Transport , Sequence Alignment , Sequence Homology , ras Proteins/genetics , ras Proteins/metabolismABSTRACT
In response to nutrient limitations, Bacillus subtilis cells undergo a series of morphological and genetic changes that culminate in the formation of endospores. Conversely, excess catabolites inhibit sporulation. It has been demonstrated previously that excess catabolites caused a decrease in culture medium pH in a process that required functional AbrB. Culture medium acidification was also shown to inhibit sigmaH-dependent sporulation gene expression. The studies reported here investigate the effects of AbrB-mediated pH sensing on B. subtilis developmental competence. We have found that neither addition of a pH stabilizer, MOPS (pH 7.5), nor null mutations in abrB blocked catabolite repression of sporulation. Moreover, catabolite-induced culture medium acidification was observed in cultures of catabolite-resistant sporulation mutants, crsA47, rvtA11, and hpr-16, despite their efficient sporulation. These results suggest that AbrB-mediated pH sensing is not the only mechanism regulating catabolite repression of sporulation. The AbrB pathway may function to channel cells toward genetic competence, as opposed to other postexponential differentiation pathways.
