Tunicamycin-resistant mutants of Bacillus amyloliquefaciens are deficient in amylase, protease and penicillinase synthesis and have altered sensitivity to antibiotics and autolysis

Mutants of Bacillus amyloliquefaciens resistant to at least 10 micrograms/ml of tunicamycin were isolated and shown to be pleiotropic. The mutants were more resistant to streptomycin, chloramphenicol, kanamycin and neomycin than was the parent strain but less resistant to penicillin G and tetracycline. They were more autolytic, presumably due to an altered cell wall. The mutants produced reduced levels of amylase, penicillinase and both metal and serine protease besides having an enhanced sporulation frequency and being more motile.     

Estimate of mean tissue O2 consumption at onset of exercise in males

A mathematical model has been developed that permitted the calculation of the flow-weighted mean tissue O2 consumption (VO2T) at the onset of a step increase in work rate. From breath-by-breath measurements of alveolar O2 consumption (VO2A) and cardiac output (Q) by impedance cardiography and assumptions about the site of depletion of O2 stores, the rate of change in O2 stores (VO2s) was determined. The sum of VO2A + VO2s = VO2T. Six very fit males performed six repetitions of each of two step increases in work rate. STlo was a transition from rest to 100-W cycling; SThi was a transition from 100- to 200-W cycling. For each work rate transition, the responses of VO2A and Q were averaged over the six repetitions of each subject and the model was solved to yield VO2T. The responses of VO2A, VO2T, and Q after the increase in work rate were fit with a monoexponential function. This function included a time constant and time delay, the sum of which gave the mean response time (MRT). In the STlo test, the MRT of VO2A (24.9 +/- 1.1 s, mean +/- SE) was longer than that of VO2T (15.3 +/- 1.3 s) and of Q (16.5 +/- 6.5 s) (P less than 0.05). The MRT of VO2T and Q did not differ significantly. Also for SThi, the MRT of VO2A (34.4 +/- 3.3 s) was significantly longer than that of VO2T (30.0 +/- 3.4 s) (P less than 0.05). The MRT of VO2T and Q (30.3 +/- 5.5 s) were not significantly different at this work rate either.(ABSTRACT TRUNCATED AT 250 WORDS)     

Demonstration that a chemically synthesized BPV1 oncoprotein and its C-terminal domain function to induce cellular DNA synthesis

Bovine papillomavirus type 1 contains the smallest known oncogene (ORF E5), encoding a hydrophobic 44 amino acid protein. To study the biochemical functions of the E5 oncoprotein, we have chemically synthesized it and several deletion mutant peptides. We demonstrate induction of cellular DNA synthesis in growth-arrested cells by microinjection of E5 oncoprotein. This activity can be broken down into two functionally distinguishable domains. Remarkably, the first domain, which alone is sufficient to induce cellular DNA synthesis, contains only the C-terminal 13 amino acids. This is the smallest known protein fragment that can autonomously activate cellular DNA synthesis. The second domain is the hydrophobic middle region, which by itself fails to induce cellular DNA synthesis but confers a 1000-fold increase in specific activity. The N-terminal one-third of the molecule is dispensable for induction of DNA synthesis.     

Hight performance gel permeation chromatography of proteins

Proteins in the molecular weight range of 10 000–170 000 were separated by high performance gel permeation chromatography. Silica particles with 30 nm or 50 nm pores were derivatized with glycidoxy-propyltrimethoxysilane and used as support. The proteins were eluted with 50% formic acid. A protein fraction which induces endodermal and mesodermal tissues in amphibian gastrula ectoderm was purified by this method.     

On the Robustness of the Geometrical Model for Cell Wall Deposition

All plant cells are provided with the necessary rigidity to withstand the turgor by an exterior cell wall. This wall is composed of long crystalline cellulose microfibrils embedded in a matrix of other polysaccharides. The cellulose microfibrils are deposited by mobile membrane bound protein complexes in remarkably ordered lamellar textures. The mechanism by which these ordered textures arise, however, is still under debate. The geometrical model for cell wall deposition proposed by Emons and Mulder (Proc. Natl. Acad. Sci. 95, 7215–7219, 1998) provides a detailed approach to the case of cell wall deposition in non-growing cells, where there is no evidence for the direct influence of other cellular components such as microtubules. The model successfully reproduces even the so-called helicoidal wall; the most intricate texture observed. However, a number of simplifying assumptions were made in the original calculations. The present work addresses the issue of the robustness of the model to relaxation of these assumptions, by considering whether the helicoidal solutions survive when three aspects of the model are varied. These are: (i) the shape of the insertion domain, (ii) the distribution of lifetimes of individual CSCs, and (iii) fluctuations and overcrowding. Although details of the solutions do change, we find that in all cases the overall character of the helicoidal solutions is preserved.     

DNA sequence plays a major role in determining nucleosome positions in yeast CUP1 chromatin.

The role of DNA sequence in determining nucleosome positions in vivo was investigated by comparing the positions adopted by nucleosomes reconstituted on a yeast plasmid in vitro using purified core histones with those in native chromatin containing the same DNA, described previously. Nucleosomes were reconstituted on a 2.5 kilobase pair DNA sequence containing the yeast TRP1ARS1 plasmid with CUP1 as an insert (TAC-DNA). Multiple, alternative, overlapping nucleosome positions were mapped on TAC-DNA. For the 58 positioned nucleosomes identified, the relative positioning strengths and the stabilities to salt and temperature were determined. These positions were, with a few exceptions, identical to those observed in native, remodeled TAC chromatin containing an activated CUP1 gene. Only some of these positions are utilized in native, unremodeled chromatin. These observations suggest that DNA sequence is likely to play a very important role in positioning nucleosomes in vivo. We suggest that events occurring in yeast CUP1 chromatin determine which positions are occupied in vivo and when they are occupied.     

The Teorell membrane oscillator as a mechano-electric transducer

Summary The results of a recent quantitative analysis of the Teorell membrane oscillator are utilized to explore its role as an excitability analogue. Special attention is paid to its role as a mechano-electric transducer. A membrane of exceptionally well-defined pore structure has been used in this study. The analogue properties arise from nonlinear coupling between water and salt fluxes. When the membrane is simultaneously subjected to controlled gradients of hydrostatic pressure, electrical potential and concentration, bi-stable stationary states can be produced. These arise from the opposing effects of pressure and electro-osmosis on the volume flow. Transitions between these states show hysteresis. The factors governing such transitions are analogous to certain types of stimuli encountered in the natural excitation process. The membrane system also shows oscillatory behavior when the hydrostatic pressure gradient is allowed to vary under constant current conditions. This property is related to the bi-stable stationary state phenomena and is compared to the regenerative behavior found in biologically excitable tissues. Particular emphasis is placed upon analogies between the membrane oscillator and certain natural tissues. The importance of the nonlinear nature of the force-flux coupling in the analogue is stressed, and its possible relevance to biological excitability indicated. Some consideration is also given to the role of electro-osmotic flux coupling in biological tissues.