Allosteric regulation of the primase (DnaG) activity by the clamp-loader (τ) in vitro

During DNA replication the helicase (DnaB) recruits the primase (DnaG) in the replisome to initiate the polymerization of new DNA strands. DnaB is attached to the τ subunit of the clamp-loader that loads the β clamp and interconnects the core polymerases on the leading and lagging strands. The τ–DnaB−DnaG ternary complex is at the heart of the replisome and its function is likely to be modulated by a complex network of allosteric interactions. Using a stable ternary complex comprising the primase and helicase from Geobacillus stearothermophilus and the τ subunit of the clamp-loader from Bacillus subtilis we show that changes in the DnaB–τ interaction can stimulate allosterically primer synthesis by DnaG in vitro. The A550V τ mutant stimulates the primase activity more efficiently than the native protein. Truncation of the last 18 C-terminal residues of τ elicits a DnaG-stimulatory effect in vitro that appears to be suppressed in the native τ protein. Thus changes in the τ–DnaB interaction allosterically affect primer synthesis. Although these C-terminal residues of τ are not involved directly in the interaction with DnaB, they may act as a functional gateway for regulation of primer synthesis by τ-interacting components of the replisome through the τ–DnaB−DnaG pathway.

Impact of Bottom Flange Confinement Reinforcement on Performance of Prestressed Concrete Bridge Girders

For many years AASHTO provided no recommendation to state DOT’s on bottom flange confinement reinforcement for their bridge superstructures. The 1996 edition of AASHTO Standard Specification for Highway Bridges stated that nominal reinforcement be placed to enclose the prestressing steel from the end of the girder for at least a distance equal to the girder’s height. A few years later the 2004 AASHTO LRFD Bridge Design Specification changed the distance over which the confinement was to be distributed from 1.0h to 1.5h, and gave minimum requirements for the amount of steel to be used, No.3 bars, and their maximum spacing, not to exceed 6”. Research was undertaken to study what impact, if any, confinement reinforcement has on the performance of prestressed concrete bridge girders. Of particular interest was the effect confinement had on the transfer length, development length, and vertical shear capacity of the fore mentioned members. First, an analytical investigation was performed on the subject, and then an experimental investigation followed which consisted of designing, fabricating, and testing eight tee-girders and three NU1100 girders with particular attention paid to the amount and distribution of confinement reinforcement placed at the end of each girder. The results of the study show: 1) neither the amount or distribution of confinement reinforcement had a significant effect on the initial or final transfer length of the prestress strands; 2) at the AASHTO calculated development length, no significant impact from confinement was found on either the nominal flexural capacity of bridge girders or bond capacity of the prestressing steel; 3) the effects from varied confinement reinforcement on the shear resistance of girders tested was negligible, however, distribution of confinement did show to have an impact on the prestressed strands’ bond capacity; 4) confinement distribution across the entire girder did increase ductility and reduced cracking under extreme loading conditions.

Deterministic Solutions for a Step-growth Polymerization

Chain topology, including branch node, chain link and cross-link dynamics that contribute to the number of elastically active strands and junctions, are calculated using purely deterministic derivations. Solutions are not coupled to population density distributions. An eigenzeit transformation assists in the conversion of expressions derived by chemical reaction principles from time to conversion space, yielding transport phenomena type expressions where the rate of change in the molar concentrations of branch nodes with respect to conversion is expressed as functions of the fraction of reactive sites on precursors and reactants. Analogies are hypothesized to exist in cross-linking space that effectively distribute branch nodes with i reacted moieties between cross-links having j bonds extending to the gel. To obtain solutions, reacted sites on nodes or links with finite chain extensions are examined in terms of stoichiometry associated with covalent bonding. Solutions replicate published results based on Miller and Macosko’s recursive procedure and results obtained from truncated weighted sums of population density distributions as suggested by Flory.