Peptide ligands that bind selectively to spores of Bacillus subtilis and closely related species

As part of an effort to develop detectors for selected species of bacterial spores, we screened phage display peptide libraries for 7- and 12-mer peptides that bind tightly to spores of Bacillus subtilis. All of the peptides isolated contained the sequence Asn-His-Phe-Leu at the amino terminus and exhibited clear preferences for other amino acids, especially Pro, at positions 5 to 7. We demonstrated that the sequence Asn-His-Phe-Leu-Pro (but not Asn-His-Phe-Leu) was sufficient for tight spore binding. We observed equal 7-mer peptide binding to spores of B. subtilis and its most closely related species, Bacillus amyloliquefaciens, and slightly weaker binding to spores of the closely related species Bacillus globigii. These three species comprise one branch on the Bacillus phylogenetic tree. We did not detect peptide binding to spores of several Bacillus species located on adjacent and nearby branches of the phylogenetic tree nor to vegetative cells of B. subtilis. The sequence Asn-His-Phe-Leu-Pro was used to identify B. subtilis proteins that may employ this peptide for docking to the outer surface of the forespore during spore coat assembly and/or maturation. One such protein, SpsC, appears to be involved in the synthesis of polysaccharide on the spore coat. SpsC contains the Asn-His-Phe-Leu-Pro sequence at positions 6 to 10, and the first five residues of SpsC apparently must be removed to allow spore binding. Finally, we discuss the use of peptide ligands for bacterial detection and the use of short peptide sequences for targeting proteins during spore formation.     

Astrobiology and Biological Chirality

The emerging discipline of astrobiology could gain valuable support from research dealing with the problems of biological chirality. The most profitable fields of common interest are: (a) living organisms under extraterrestrial conditions, (b) extraterrestrial signatures of life and (c) origin(s) of biological chirality. These areas of complementary and overlapping fields are analysed on the basis of selected references. Presented at: National Workshop on Astrobiology: Search for Life in the Solar System, Capri, Italy, 26 to 28 October, 2005.     

Synthesis of oligopeptides with the sequence SXWS and their chemotactic effects on a ciliated protozoan Tetrahymena pyriformis.

In this paper, the solid phase synthesis and chemical characterization of members of an SXWS sub-library (SAWS, SDWS and SKWS) as well as the comparison of their chemotactic properties with those of SEWS, which exhibits a prominent effect at 10(-12) M on a ciliated protozoan, Tetrahymena pyriformis, are described. We found that the chemotaxis of cells induced with the SXWS peptides varied according to the nature of the amino acid residue (Ala, Asp, Lys) in position X. The chemotactic activity of SEWS was not surpassed by any of three new tetrapeptides, although SAWS was also chemoattractant. Interestingly, SDWS, with an acidic side chain at position X, could not elicit any chemotactic response. SKWS, however, showed mild but significant chemorepellent activity over a wide concentration range. Chemotactic selection studies showed that the two chemoattractant peptides (SAWS and SEWS) had an expressed ability to select high-responder offspring cell populations. Peptides with neutral (SDWS) or chemorepellent (SKWS) properties were not able to select such subpopulations from the mixed cultures of Tetrahymena, indicating that the chemotactic response elicited by SXWS peptides is ligand-specific. For ligand-binding experiments N-terminally labelled fluorescent derivatives of SXWS peptides were prepared. applying [4-[7-hydroxycoumaryl]]acetic acid (Hca-OH) or 4-ethoxymethylene-2-[1]-naphthyl-5(4H)-oxazolone (naOx-OEt) as markers. Hca-OH was introduced using an active ester technique as the last step of SPPS, or after cleavage in solution. The oxazolone naOx-OEt reacted with the amino group of the peptide by liberation of EtOH. The binding characteristics of fixed Tetrahymena cells with the naOx-labelled peptides showed good correlation between binding profiles and chemotactic responsiveness (SEWS > SAWS > SDWS - SKWS). A similar binding pattern was observed in the case of Hca-peptides (SEWS > SAWS > SDWS). Hca-SKWS, however, bound remarkably to the cell surface. The binding activity of the Hca-peptides was less pronounced than that of the naOx-peptides, indicating the importance of the fluorophores applied.     

Autophagy-dependent survival is controlled with a unique regulatory network upon various cellular stress events

Although autophagy is a type of programmed cell death, it is also essential for cell survival upon tolerable level of various stress events. For the cell to respond adequately to an external and/or internal stimulus induced by cellular stress, autophagy must be controlled in a highly regulated manner. By using systems biology techniques, here we explore the dynamical features of autophagy induction. We propose that the switch-like characteristic of autophagy induction is achieved by a control network, containing essential feedback loops of four components, so-called autophagy inducer, autophagy controller, mTORC1 and autophagy executor, respectively. We show how an autophagy inducer is capable to turn on autophagy in a cellular stress-specific way. The autophagy controller acts as a molecular switch and not only promotes autophagy but also blocks the permanent hyperactivation of the process via downregulating the autophagy inducer. In this theoretical analysis, we explore in detail the properties of all four proposed controlling elements and their connections. Here we also prove that the kinetic features of this control network can be considered accurate in various stress processes (such as starvation, endoplasmic reticulum stress and oxidative stress), even if the exact components may be different. The robust response of the resulting control network is essential during cellular stress.Subject terms: Biochemistry, Molecular biology     

An aggregation attractant for the sugar-beet weevil, Bothynoderes punctiventris

During our screening studies, attractiveness of a ternary mixture of synthetic Grandlure I [racemic cis‐1‐methyl‐2‐(1‐methylenethenyl)‐cyclobutane ethanol], Grandlure II [(Z)‐2‐(3,3‐dimethyl)cyclohexylidene ethanol], and Grandlure III–IV [(Z)‐ and (E)‐2‐ochtodenal; (Z)‐ and (E)‐(3,3‐dimethyl)cyclohexylidene acetaldehyde] for the sugar‐beet weevil, Bothynoderes punctiventris Germar (Coleoptera: Curculionidae), was observed in field‐trapping tests at several sites in Hungary and Serbia. The mixture attracted both males and females. Later tests revealed that of the components in the ternary mixture, only Grandlure III–IV were responsible for attraction, and the addition of Grandlures I or II in varying percentages had no influence on trap captures. Traps baited with 50–50 000 µg of Grandlure III–IV on rubber or polyethylene dispensers yielded high catches of weevils. When testing synthetic samples enriched in the respective geometrical isomer, Grandlure IV had a tendency of catching more weevils, but differences were not significant from lower catches by a 1:1 Z:E blend or Grandlure III. In gas chromatography–flame ionization detection/electroantennographic detection studies, antennae of both female and male weevils were more responsive to the (E)‐ than to the (Z)‐isomer suggesting a more important role for Grandlure IV. Efforts to verify the presence of Grandlure III or IV in volatiles collected from either sex of live sugar‐beet weevils or body washings with pentane remained inconclusive. Traps baited with Grandlure III–IV can now be used as sensitive and powerful trapping tools in the control of the sugar‐beet weevil.     

Past and future climate-driven shifts in the distribution of a warm-adapted bird species,the European Roller Coracias garrulus

ABSTRACT

Capsule: The distribution range of the European Roller Coracias garrulus has undergone large changes over geological times, but although the species is warm-adapted, the human induced climate change is predicted to affect negatively the range of the currently large populations.

Aim: Information on species-specific vulnerability to climate change is crucial not only for designing interventions and setting conservation goals, but also to inform conservation decision-making. Our goal was to map climate suitability for the European Roller in the Western Palaearctic under current climate, and for past (last glacial maximum and mid-Holocene) and future (2050 and 2070) climate scenarios.

Methods: We used MaxEnt for species distribution modelling based on the reconstructed distribution map of the species.

Results: Our results suggest that during glacial periods Rollers persisted in small southern refugia, and then spread and colonized northern latitudes during the mid-Holocene. In the future, our models forecast a shift in climatically suitable range towards northern latitudes and an overall small range contraction (4.5–5.5%). Warmer temperatures will increase climate suitability in northern countries where the species is currently declining or became locally extinct. On the other hand, wide suitable areas under current climatic conditions are predicted to become unsuitable in the future (35–38% by 2050 and 2070, respectively), significantly impacting large populations such as those in Romania, Spain, Bulgaria and Hungary. French and Italian populations are identified to be future key populations for Roller conservation.

Conclusions: Our findings suggest that future climate changes will likely amplify the impacts of existing threats on the majority of large European Roller populations in Europe.