The Polypeptide Transport-associated (POTRA) Domains of TpsB Transporters Determine the System Specificity of Two-partner Secretion Systems

The two-partner secretion (TPS) systems of Gram-negative bacteria secrete large TpsA exoproteins by a dedicated TpsB transporter in the outer membrane. TpsBs contain an N-terminal module located in the periplasm that includes two polypeptide transport-associated (POTRA) domains. These are thought to initiate secretion of a TpsA by binding its N-terminal secretion signal, called the TPS domain. Neisseria meningitidis encodes up to five TpsA proteins that are secreted via only two TpsB transporters: TpsB1 and TpsB2. Of these two, the TpsB2 recognizes the TPS domains of all TpsAs, despite their sequence diversity. By contrast, the TpsB1 shows a limited recognition of a TPS domain that is shared by two TpsAs. The difference in substrate specificity of the TpsBs enabled us to investigate the role of the POTRA domains in the selection of TPS domains. We tested secretion of TPS domains or full-length TpsAs by TpsB mutants with deleted, duplicated, and exchanged POTRA domains. Exchanging the two POTRA domains of a TpsB resulted in a switch in specificity. Furthermore, exchanging a single POTRA domain showed that each of the two domains contributed to the cargo selection. Remarkably, the order of the POTRA domains could be reversed without affecting substrate selection, but this aberrant order did result in an alternatively processed secretion product. Our results suggest that secretion of a TpsA is initiated by engaging both POTRA domains of a TpsB transporter and that these select the cognate TpsAs for secretion.     

Response of bacterial communities to cyanobacterial harmful algal blooms in Lake Taihu,China

Cyanobacterial harmful algal blooms are prevalent around the world, influencing aquatic organisms and altering the physico-chemical properties in freshwater systems. However, the response of bacterial communities to toxic cyanobacterial blooms and associated microcystins (MC) remain poorly understood even though global concentrations of MC have increased dramatically in the past few decades. To address this issue, the dynamics of bacterial community composition (BCC) in the water column and how BCC is influenced by both harmful cyanobacterial blooms and environmental factors were investigated on a monthly basis from August 2013 to July 2014 in Lake Taihu, China. Non-metric multidimensional scaling (NMDS) revealed that seasonal variation in BCC was significant, and that the succession of BCC greatly depends on changes in environmental conditions. Redundancy analysis (RDA) results showed that the overall variation of BCC was explained mainly by dissolved oxygen (DO), nitrate nitrogen (NO₃⁻-N), and Microcystis. The alpha biodiversity of the bacterial community was different among months with the highest diversity in February and the lowest diversity in October. Furthermore, significant negative relationships were found between alpha biodiversity indices and Microcystis abundance as well as with intracellular MC concentrations, indicating that Microcystis and associated MC may influence the bacterial community structure by reducing its biodiversity. This study shows that potential associations exist between toxic cyanobacterial blooms and bacterial communities but more investigations are needed to obtain a mechanistic understanding of their complex relationships.     

Structural Basis for Intrinsic Thermosensing by the Master Virulence Regulator RovA of Yersinia

Pathogens often rely on thermosensing to adjust virulence gene expression. In yersiniae, important virulence-associated traits are under the control of the master regulator RovA, which uses a built-in thermosensor to control its activity. Thermal upshifts encountered upon host entry induce conformational changes in the RovA dimer that attenuate DNA binding and render the protein more susceptible to proteolysis. Here, we report the crystal structure of RovA in the free and DNA-bound forms and provide evidence that thermo-induced loss of RovA activity is promoted mainly by a thermosensing loop in the dimerization domain and residues in the adjacent C-terminal helix. These determinants allow partial unfolding of the regulator upon an upshift to 37 °C. This structural distortion is transmitted to the flexible DNA-binding domain of RovA. RovA contacts mainly the DNA backbone in a low-affinity binding mode, which allows the immediate release of RovA from its operator sites. We also show that SlyA, a close homolog of RovA from Salmonella with a very similar structure, is not a thermosensor and remains active and stable at 37 °C. Strikingly, changes in only three amino acids, reflecting evolutionary replacements in SlyA, result in a complete loss of the thermosensing properties of RovA and prevent degradation. In conclusion, only minor alterations can transform a thermotolerant regulator into a thermosensor that allows adjustment of virulence and fitness determinants to their thermal environment.     

Effect of bacterial heterogeneity on adhesion to uniform collectors by monoclonal populations

Abstract Transport of bacteria over significant distances through aquifer sediments occurs primarily among bacteria with low affinity for sediment materials. Bacterial affinity for a uniform collector surface has been represented quantitatively by a collision efficiency (α), defined as the fraction of colliding cells that adhere to the collector surface. Using a new method for estimating α during advective transport of monoclonal bacterial populations through a uniform bed of 40-μm borosilicate glass spheres, we found that α decreased 10-fold over a bed depth of only 1 cm. Depth-dependent differences in α were not related to variation in bacterial size or intra-strain genetic variation. Intra-population heterogeneity in biocolloid-collector affinity may be important determinant of subsurface bacterial transport characteristics, with critical implications for pathogen transport and dispersal of bacteria for the remediation of hazardous waste.     

Quantitative genetic analysis of biotypes of the brown planthopper Nilaparvata lugens: heritability of virulence to resistant rice varieties

The virulence of Nilaparvata lugens Stål (Homoptera: Delphacidae) to a resistant variety of rice, Oryza sativa L., is suggested to be under polygenic control. To determine whether the virulence of N. lugens is a quantitative character that has continuous distribution or a threshold character that has a few discontinuous phenotypic forms and is determined by some underlying continuous variable, I examined the frequency distribution of honeydew excretion that has been used as a measure of ability of attacking the rice plants, and the relationship between honeydew excretion and the oviposition rate of N. lugens females using a japonica rice line Saikai 190 which has a resistance gene Bph 1. The frequency distribution in honeydew excretion significantly differed from the normal distribution, but two distributions below and above 10 mg honeydew excretion for two days did not significantly deviate from normality, suggesting a bimodal distribution. There were significant differences in the proportion of females ovipositing and the number of eggs deposited between the female groups that excreted 0–10 mg honeydew and more than 20 mg honeydew. Within these female groups, however, the reproductive performances were not different. Thus, virulence of N. lugens can be analyzed as a threshold character that has two distinct phenotypes, virulent and avirulent. I estimated the heritabilities of the virulence of N. lugens by parent-offspring regression using the percentage of virulent females in a full-sib family. The estimates of heritability were 0.41 on Saikai 190 and 0.55 on ASD7, carrying the bph 2 gene. These results show that the N. lugens population has substantial genetic variation in virulence. The regression coefficients on female and male parents were similar, suggesting similar genetic contributions by both parents. When the parental families were examined on Saikai 190 and the progeny on ASD7, the regression coefficient approached zero. There may be a low genetic correlation between virulences to the two varieties.