Analysis of the motA flagellar motor gene from Rhodobacter sphaeroides a bacterium with a unidirectional, stop-start flagellum

Rhodobacter sphaeroides swims by unidirectional rotation of a single medial flagellum, re-orienting randomly by Brownian motion when flagellar rotation tops and restarts. Previously we identified a mutant with a paralysed flagellum, which was complemented by a Rhodobacter gene that had homology to motB of Escherichia coli , a bacterium with bidirectional flagella. In the current work, interposon mutagenesis upstream of the Rhodobacter motB gene gave rise to another paralysed mutant, RED5. DNA sequence analysis of this upstream region showed one open reading frame, the predicted polypeptide sequence of which shows homology to the MotA protein of E. coli . MotA is thought to be a proton 'pore' involved in converting proton-motive force into flagellar rotation. Several potential proton-binding amino acids were conserved between putative membrane-spanning regions of R. sphaeroides and E. coli MotA sequences, along with a highly charged cytoplasmic linker region. Complementation studies with mutant RED5 showed the presence of an active promoter upstream from motA which was found to be necessary for expression of both motA and motB , Examination of the upstream DNA sequence showed only one putative promoter-like sequence which resembled a σ⁵⁴- type promoter, including a potential enhancer binding site. The overall similarities between the R. sphaeroides MotA protein and those from other bacteria suggest that, despite the novel unidirectional rotation of he R. sphaeroides flagellum, the function of the MotA protein is similar to that in bacteria with bidirectional flagella.     

One short well conserved region of Alu-sequences is involved in human gene rearrangements and has homology with prokaryotic chi.

Alu elements have repeatedly been found involved in gene rearrangements in humans. Although these elements have been suggested to stimulate gene rearrangements, sparse information is available for the possible mechanism(s) of these events. Here we present a compilation of Alu elements that have been involved in recombinational events leading to gene rearrangements, indicating the presence of a common 26 bp core sequence at or close to the sites of recombination. Besides the obvious possibility of retrotransposition, gene rearrangements may be induced by sequences that stimulate genetic recombination. We suggest that the core sequence stimulates recombination and may thereby cause the frequent involvement of these elements in gene rearrangements. Curiously, the core sequence contains the pentanucleotide motif CCAGC, which is also part of chi, an 8 bp sequence known to stimulate recBC mediated recombination in Escherichia coli.     

Characterization of eukaryotic-like kinase activity in Escherichia coli using the gene-protein database

Abstract The gene-protein database was used to obtain the two-dimensional polyacrylamide gel coordinates of proteins phosphorylated in extracts of Escherichia coli including those phosphorylated by eukaryotic-like kinase activities. These suggest that the phosphoproteins correspond to, or co-migrate with, the product of an open reading frame at 1.3 min (Orf80), Enzyme 1 of the phosphoenolpyruvate-dependent phosphotransferase system (Ptsl), the tRNA synthetase for histidine (HisS), and proteins involved in the response to carbon starvation and quinone treatment.     

Malignant transformation of rat thyroid cells transfected with the human TSH receptor cDNA.

Growth and function of well differentiated FRTL-5 thyroid cells depend on thyrotropin as its main regulatory hormone. We demonstrate here that stable transfection of FRTL-5 cells with the human thyrotropin receptor cDNA results in cellular transformation of these cells with altered cell shape and loss of contact inhibition. The transformed cells replicate in soft agar and form invasive tumors when cell suspensions are implanted onto nude mice. They have lost their thyrotropin dependent growth and their ability to concentrate iodide and synthesize thyroglobulin. But they still express the rat thyrotropin receptor mRNA and accumulate cAMP in response to thyrotropin stimulation. However, although the full length human thyrotropin receptor cDNA is integrated into their genome, transformed cells do not express the human thyrotropin receptor mRNA.     

Characterization of the pH-mediated solubility of Bacillus thuringiensis var. san diego native delta-endotoxin crystals.

Native crystals of Bacillus thuringiensis var. san diego, a coleopteran-specific delta-endotoxin, were metabolically labelled with [35S]methionine. Specific activity was 82,000 CPM/micrograms (2.44 Ci/mmol). Using a universal buffer formulated with the same ionic strength at every pH, we determined that native crystals dissolve above pH 10 and below pH 4. At the acidic pH, the rate of solubilization was substantially slower than at the alkaline pH. Recrystallization rates for the toxin were similar regardless of solubilization conditions. The banding patterns in denatured polyacrylamide gel electrophoresis were unaffected by solubilization conditions. Toxicity was higher for soluble toxin compared to crystal toxin, but virtually identical for the acidic and alkaline produced solutions. Acid solubilization is significant because of the acidic midgut of susceptible Coleoptera.     

Modulation of transient type K channel cloned from rat heart.

We have cloned a transient type K channel from rat heart (RH10) and coexpressed a metabotropic glutamate receptor (mGluR5) to study the functional modulation of RH10 coupled to the phosphatidylinositol (PI) hydrolysis. Stimulation of mGluR5 suppressed peak amplitude of RH10 current and affected voltage dependence of activation and inactivation of the channel.     

Introduction to the Symposium: What is Computational Neurobiology?

SYNOPSIS. Current advances in computational biology are derivedfrom two sets of ideas established in the work of Santiago Ramóny Cajal. One is the neuron doctrine, which holds that neuronsare the functional units of the nervous system, and has ledto detailed models of neuronal properties based on increasinginformation on the physical and chemical properties of neurons.The second is the idea of networks of neurons with specificpatterns of interconnections that has led to a variety of mathematicalmethods of analyzing such networks. Future work in computationalneurobiology promises to be a blend of these two modeling approaches.