Arieh Zaritsky

Complete sequence and organization of pBtoxis, the toxin-coding plasmid of Bacillus thuringiensis subsp. israelensis

ABSTRACT The entire 127,923-bp sequence of the toxin-encoding plasmid pBtoxis from Bacillus thuringiensis subsp. israelensis is presented and analyzed. In addition to the four known Cry and two known Cyt toxins, a third Cyt-type sequence was found with an additional C-terminal domain previously unseen in such proteins. Many plasmid-encoded genes could be involved in several functions other than toxin production. The most striking of these are several genes potentially affecting host sporulation and germination and a set of genes for the production and export of a peptide antibiotic.

On microbial states of growth.

It is crucial to the reproducibility of results and their proper interpretation that the conditions under which experiments are carried out be defined with rigour and consistency, in this review we attempt to clarify the differences and interrelationships among steady, balanced and exponential states of culture growth. Basic thermodynamic concepts are used to introduce the idea of steady‐state growth in open, biological systems. The classical, sometimes conflicting, definitions of steady‐state and balanced growth are presented, and a consistent terminology is proposed. The conditions under which a culture in balanced growth is also in exponential growth and in steady‐state growth are indicated. It is pointed out that steady‐state growth always implies both balanced and exponential growth, and examples in which the converse does not hold are described. More complex situations are then characterized and the terminology extended accordingly. This leads to the notion of normal growth and growth that can be synchronous or otherwise unbalanced but still reproducible, and to the condition of approximate steady state manifested by growth in batch culture and by asymmetrically dividing cells, which is analysed in some detail.

A UV Tolerant Mutant of Bacillus thuringiensis subsp. kurstaki Producing Melanin

A UV-resistant mutant (Bt-m) of Bacillus thuringiensis subsp. kurstaki, producing a dark brown pigment, identified as melanin, was studied. Bt-m had higher larvicidity against Heliothis armigera than its parent. Survival of Bt-m spores and their insecticidal activity to irradiation at 254 nm and 366 nm were higher than those of the parent. The only toxic polypeptide produced by Bt-m was Cry1Ac (130 kDa); it lost cry1Aa, cry2Aa, and cry2Ab.

Transcription‐ and translation‐dependent changes in membrane dynamics in bacteria: testing the transertion model for domain formation

Cell cycle events have been proposed to be triggered by the formation of membrane domains in the process of coupled transcription, translation and insertion (‘transertion’) of nascent membrane and exported proteins. Disruption of domain structure should lead to changes in membrane dynamics. Membrane viscosity of Escherichia coli and Bacillus subtilis decreased after inhibition of protein synthesis by chloramphenicol or puromycin, or of RNA initiation by rifampicin, but not after inhibition of RNA elongation by streptolydigin or amino acid starvation of a stringent strain. The decrease caused by inhibitors of protein synthesis was prevented by streptolydigin if added simultaneously, but was not reversed if added later. The drug‐induced decrease in membrane viscosity is energy dependent: it did not happen in KCN‐treated cells. All treatments decreasing membrane viscosity also induced nucleoid compaction and fusion. Inhibition of macromolecular synthesis without membrane perturbation caused nucleoids to expand. Changes in membrane dynamics were also displayed during a nutritional shift‐down transition that causes imbalance in macromolecular syntheses. The results are consistent with the transertion model, predicting dissipation of membrane domains by termination of protein synthesis or detachment of polysomes from DNA; domain structure is conserved if the transertion process is ‘frozen’.

Richness and Diversity in Dust Stormborne Biomes at the Southeast Mediterranean

Dust storms include particulate matter that is transported over land and sea with biota that could impact downwind ecosystems. In addition to the physico-chemical compositions, organismal diversities of dust from two storm events in southern Israel, December 2012 (Ev12) and January 2013 (Ev13), were determined by pyro-sequencing using primers universal to 16S and 18S rRNA genes and compared. The bio-assemblages in the collected dust samples were affiliated with scores of different taxa. Distinct patterns of richness and diversity of the two events were influenced by the origins of the air masses: Ev13 was rich with reads affiliated to Betaproteobacteria and Embryophyta, consistent with a European origin. Ev12, originated in north-Africa, contained significantly more of the Actinobacteria and fungi, without conifers. The abundance of bacterial and eukaryotic reads demonstrates dissemination of biological material in dust that may impose health hazards of pathogens and allergens, and influence vegetation migration throughout the world.

High-resolution crystal structure of activated Cyt2Ba monomer from Bacillus thuringiensis subsp. israelensis.

The Cyt family of proteins consists of delta-endotoxins expressed during sporulation of several subspecies of Bacillus thuringiensis. Its members possess insecticidal, hemolytic, and cytolytic activities through pore formation and attract attention due to their potential use as vehicles for targeted membrane destruction. The delta-endotoxins of subsp. israelensis include three Cyt species: a major Cyt1Aa and two minor proteins, Cyt2Ba and Cyt1Ca. A cleaved Cyt protein that lacks the N- and C-terminal segments forms a toxic monomer. Here, we describe the crystal structure of Cyt2Ba, cleaved at its amino and carboxy termini by bacterial endogenous protease(s). Overall, its fold resembles that of the previously described volvatoxin A2 and the nontoxic form of Cyt2Aa. The structural similarity between these three proteins may provide information regarding the mechanism(s) of membrane-perforating toxins.

Bacterial debris—an ecological mechanism for coexistence of bacteria and their viruses

A model of bacteria and phage survival is developed based on the idea of shielding by bacterial debris in the system. This model is mathematically formulated by a set of four nonlinear difference equations for susceptible bacteria, contaminated bacteria, bacterial debris and phages. Simulation results show the possibility of survival, and domains of existence of stable and unstable solutions

Dimensional rearrangement of rod-shaped bacteria following nutritional shift-up. II. Experiments with Escherichia coliBr☆

The dimensions of Escherichia coliBr (strain H266) in transition between two states of balanced growth, were determined from electron micrographs of fixed cells by sampling the culture at various times following nutritional shift-up from a doubling time of 72 min to one of 24 min. Mean cell length rises immediately and overshoots its final steady-state value, cell diameter increases monotonically; both approach their asymptotic levels only after several hours. The results are compared with the dimensions predicted by each of two models of cell growth and morphogenesis in rod-shaped bacteria. The first attributes cell elongation to circular zones that double in number at a particular time during the cell cycle and which act at rates proportional to the growth rate; the second is similar, except that it considers surface growth rather than length extension as the active process, length being determined passively. Two possibilities are examined, that the zonal growth rate adjusts immediately to the new growth conditions, and that it does so gradually. The experimental data appear consistent with the gradual response version of the surface growth model.

On dimensional determination of rod-shaped bacteria.

A formal model which describes quantitatively how the dimensions of rod-shaped bacteria are determined is presented. It is proposed that the rate of extension in length is proportional to the number of chromosome termini and the diameter to the number of chromosome replication positions, and data in the literature are compared with the theoretical predictions made.

Comparative Sensitivity to UV-B Radiation of Two Bacillus thuringiensis Subspecies and Other Bacillus sp.

Susceptibility of Bacillus thuringiensis spores and toxins to the UV-B range (280–330 nm) of the solar spectrum reaching Earths surface may be responsible for its inactivation and low persistence in nature. Spores of the mosquito larvicidal B. thuringiensis subsp. israelensis were significantly more resistant to UV-B than spores of the lepidopteran-active subsp. kurstaki. Spores of subsp. israelensis were as resistant to UV-B as spores of B. subtilis and more resistant than spores of the closely related B. cereus and another mosquito larvicidal species B. sphaericus. Sensitivity of B. thuringiensis subsp. israelensis spores to UV-B radiation depended upon their culture age; 24-h cultures, approaching maximal larvicidal activity, were still sensitive. Maximal resistance to UV-B was achieved only at 48 h.