To ensure the optimal infectivity on contact with host cells, pathogenic Pseudomonas syringae has evolved a complex mechanism to control the expression and construction of the functional type III secretion system (T3SS) that serves as a dominant pathogenicity factor. In this study, we showed that the hrpF gene of P. syringae pv. averrhoi, which is located upstream of hrpG, encodes a T3SS-dependent secreted/translocated protein. Mutation of hrpF leads to the loss of bacterial ability on elicitation of disease symptoms in the host and a hypersensitive response in non-host plants, and the secretion or translocation of the tested T3SS substrates into the bacterial milieu or plant cells. Moreover, overexpression of hrpF in the wild-type results in delayed HR and reduced t3ss expression. The results of protein-protein interactions demonstrate that HrpF interacts directly with HrpG and HrpA in vitro and in vivo, and protein stability assays reveal that HrpF assists HrpA stability in the bacterial cytoplasm, which is reduced by a single amino acid substitution at the 67th lysine residue of HrpF with alanine. Taken together, the data presented here suggest that HrpF has two roles in the assembly of a functional T3SS: one by acting as a negative regulator, possibly involved in the HrpSVG regulation circuit via binding to HrpG, and the other by stabilizing HrpA in the bacterial cytoplasm via HrpF-HrpA interaction prior to the secretion and formation of Hrp pilus on the bacterial surface.
We used oscillatory optical tweezers to investigate the microrheological properties of Sodium polystyrene sulfonate (NaPSS; Mw = 70 kDa) polymer solutions with different concentrations from 0.001 mM to 10 mM in terms of elastic modulus G'(ω) and loss modulus G"(ω) as a function of angular frequency (ω) in the range of 6 rad/s to 6000 rad/s. The viscoelastic properties (including zero-shear-rate viscosity, crossing frequency and transition frequency) as a function of polymer concentration, deduced from our primary data, reveal the subtle structural changes in the polymer solutions as the polymer concentration increases from dilute to semi-dilute regimes, passing through the critical micelle formation concentration and the polymer overlapping concentration. The experimental results are consistent with the Maxwell model in some regime, and with the Rouse model in other, indicating the transient network character and the micelles formation in different regimes.
In this paper, we report the viscoelastic properties of sodium polystyrene sulfonate (NaPSS) solution with different concentrations (in the range of 10 -4 M to 10 -3 M) and with different molecular weights (70 kDa vs. 200 kDa) investigated via Diffusing Wave Spectroscopy (DWS). The viscoelastic properties of the sample solutions are characterized in terms of the elastic modulus G' and the viscous modulus G" as a function of frequency (f), and also in terms of the polymer disentanglement time (τ); the effect of polymer concentration and molecular weight on these parameters are presented. Our experimental results indicate that (1) both the viscous modulus G" and the disentanglement time (τ) increase with molecular concentration, whereas the elastic modulus G' is relatively insensitive to molecular concentration, and (2) for the same concentration, all the 3 parameters (i.e., the elastic modulus G', the viscous modulus G", and disentanglement time τ) increase as the molecular weight increases.
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