Structure-Function Analysis of the HrpB2-HrcU Interaction in the Xanthomonas citri Type III Secretion System

Bacterial type III secretion systems deliver protein virulence factors to host cells. Here we characterize the interaction between HrpB2, a small protein secreted by the Xanthomonas citri subsp. citri type III secretion system, and the cytosolic domain of the inner membrane protein HrcU, a paralog of the flagellar protein FlhB. We show that a recombinant fragment corresponding to the C-terminal cytosolic domain of HrcU produced in E. coli suffers cleavage within a conserved Asn264-Pro265-Thr266-His267 (NPTH) sequence. A recombinant HrcU cytosolic domain with N264A, P265A, T266A mutations at the cleavage site (HrcUAAAH) was not cleaved and interacted with HrpB2. Furthermore, a polypeptide corresponding to the sequence following the NPTH cleavage site also interacted with HrpB2 indicating that the site for interaction is located after the NPTH site. Non-polar deletion mutants of the hrcU and hrpB2 genes resulted in a total loss of pathogenicity in susceptible citrus plants and disease symptoms could be recovered by expression of HrpB2 and HrcU from extrachromossomal plasmids. Complementation of the ΔhrcU mutant with HrcUAAAH produced canker lesions similar to those observed when complemented with wild-type HrcU. HrpB2 secretion however, was significantly reduced in the ΔhrcU mutant complemented with HrcUAAAH, suggesting that an intact and cleavable NPTH site in HrcU is necessary for total functionally of T3SS in X. citri subsp. citri. Complementation of the ΔhrpB2 X. citri subsp. citri strain with a series of hrpB2 gene mutants revealed that the highly conserved HrpB2 C-terminus is essential for T3SS-dependent development of citrus canker symptoms in planta

IMPURITY CONTENT CHARACTERIZATION OF BRAZILIAN QUARTZ LASCAS

Lattice impurities (Al, Li, Fe) free from the influence of cations in inclusion liquid were evaluated by atomic absorption spectrophotometry (AAS) in quartz lascas taken from 10 deposits with known localities in three states, Minas Gerais, Tocantins and Bahia. Levels of Al content and associated Li content depend on the localities of deposits. Quartz deposit regions with high Al content are composed of granite, gneiss, and pegmatite as surrounding rocks. It was concluded that the charge compensation for Al-Si substitution in quartz lascas was realized substantially by the formation of Al-Li and Al-OH centers which were confirmed from the linear relation of Al-Li plots by AAS data and IR spectra due to Al-OH center. The variety in Al-Li and Al-OH correlations was attributed to the fluctuation of growing conditions of quartz in nature.3071489149

Effect of particle size in the TL response of natural quartz sensitized by high dose of gamma radiation and heat-treatments

This work investigates the effect of particle size in the thermoluminescence (TL) response of a quartz crystal that was initially crushed and classified into ten size fractions between 38 μm and 5 mm. Aliquots of each size fraction were sensitized with a dose of 25 kGy of γ rays and heat-treatments at 400 °C. TL glow curves of sensitized and non-sensitized samples were recorded as a function of different test-doses of γ rays. For the non-sensitized samples, the TL peak near 325 °C increases with the decrease in particle size. In the case of sensitized samples, a strong TL peak near 300 °C increases with the increase in particle size up to mean grain size equal to 304 μm. Above 304 μm, an abrupt reduction in the TL intensity is noticed for the sensitized peak. These effects are discussed in relation to the specific surface area of quartz particles and the intensity of the electron paramagnetic resonance signal of the E'1 center induced by the sensitization process