Magnetotactic cocci swim persistently along local magnetic field lines in a preferred direction that corresponds to downward migration along geomagnetic field lines. Recently, high cell concentrations of magnetotactic cocci have been found in the water columns of chemically stratified, marine and brackish habitats, and not always in the sediments, as would be expected for persistent, downward-migrating bacteria. Here we report that cells of a pure culture of a marine magnetotactic coccus, designated strain MC-1, formed microaerophilic bands in capillary tubes and used aerotaxis to migrate to a preferred oxygen concentration in an oxygen gradient. Cells were able to swim in either direction along the local magnetic field and used magnetotaxis in conjunction with aerotaxis, i.e., magnetically assisted aerotaxis, or magneto-aerotaxis, to more efficiently migrate to and maintain position at their preferred oxygen concentration. Cells of strain MC-1 had a novel, aerotactic sensory mechanism that appeared to function as a two-way switch, rather than the temporal sensory mechanism used by other bacteria, including Magnetospirillum megnetotacticum, in aerotaxis. The cells also exhibited a response to short-wavelength light (< or = 500 nm), which caused them to swim persistently parallel to the magnetic field during illumination.

We identified a protein, Aer, as a signal transducer that senses intracellular energy levels rather than the external environment and that transduces signals for aerotaxis (taxis to oxygen) and other energy-dependent behavioral responses in Escherichia coli. Domains in Aer are similar to the signaling domain in chemotaxis receptors and the putative oxygen-sensing domain of some transcriptional activators. A putative FAD-binding site in the N-terminal domain of Aer shares a consensus sequence with the NifL, Bat, and Wc-1 signal-transducing proteins that regulate gene expression in response to redox changes, oxygen, and blue light, respectively. A double mutant deficient in aer and tsr, which codes for the serine chemoreceptor, was negative for aerotaxis, redox taxis, and glycerol taxis, each of which requires the proton motive force and͞or electron transport system for signaling. We propose that Aer and Tsr sense the proton motive force or cellular redox state and thereby integrate diverse signals that guide E. coli to environments where maximal energy is available for growth.

kenji@cryst.bioc.cam.ac.uk

The task of generating a nonredundant set of low-energy conformations for small molecules is of fundamental importance for many molecular modeling and drug-design methodologies. Several approaches to conformer generation have been published. Exhaustive searches suffer from the exponential growth of the search space with increasing degrees of conformational freedom (number of rotatable bonds). Stochastic algorithms do not suffer as much from the exponential increase of search space and provide a good coverage of the energy minima. Here, the use of a multiobjective genetic algorithm in the generation of conformer ensembles is investigated. Distance geometry is used to generate an initial conformer, which is then subject to geometric modifications encoded by the individuals of the genetic algorithm. The geometric modifications apply to torsion angles about rotatable bonds, stereochemistry of double bonds and tetrahedral chiral centers, and ring conformations. The geometric diversity of the evolving conformer ensemble is preserved by a fitness-sharing mechanism based on the root-mean-square distance of the atomic coordinates. Molecular symmetry is taken into account in the distance calculation. The geometric modifications introduce strain into the structures. The strain is relaxed using an MMFF94-like force field in a postprocessing step that also removes conformational duplicates and structures whose strain energy remains above a predefined window from the minimum energy value found in the set. The implementation, called Balloon, is available free of charge on the Internet ( http://www.abo.fi/~mivainio/balloon/).

A computer analysis of the amino acid sequences from the putative gene products of retroviral pol genes has revealed a 150-residue segment that is homologous with the ribonuclease H of Escherichia coli. The segment occurs at the carboxyl terminus of the region assigned to the 90-kDa reverse transcriptase polypeptide. In contrast, a section nearer the amino terminus of this sequence can be aligned with nonretroviral polymerases. (5), and an endonuclease ("integrase") that is essential for the integration of the newly synthesized DNA into the host genome (6).The pol gene of retroviruses is expressed initially as a gag-pol precursor that is proteolytically processed to a number of small gag proteins, an approximately 90-kDa protein encompassing both RNA-directed DNA polymerase (reverse transcriptase) and ribonuclease H activities, and, finally, a 40-kDa fragment with endonuclease activity (7). Several reports have presented evidence that the ribonuclease H activity of the 90-kDa reverse transcriptase portion is associated with the amino-terminal end of that protein, and by implication, that the DNA polymerase activity is at the carboxyl-terminal end. These conclusions are based on experiments involving deletion mutants (2), on the one hand, and antibodies to synthetic peptides modeled on the putative sequences, on the other (3).We now suggest that the opposite must be true: the ribonuclease H activity should be situated at the carboxyl terminus, and the DNA polymerase, at the amino terminus. We draw this conclusion on the basis of comparisons of the retroviral sequences with those of nonviral enzymes of similar function. In this regard, we have uncovered a significant resemblance between a 150-residue segment at the carboxyl-terminal end of the 90-kDa fragment and the reported sequence of a ribonuclease H from Escherichia coli. We also provide an alignment of a segment near the amino terminus of the 90-kDa polypeptide with highly conserved sequences from many other polymerases, including the a subunit of E. coli DNA-directed RNA polymerase. Finally, there is a distinctive sequence in the endonuclease sequence that is characteristic of a zinc-binding segment. METHODSThe sequences used were taken from the 1985 version of NEWAT (8) identical (Fig. 1). Binary comparison of each of the retroviral sequences with the E. coliribonuclease H sequence, followed by statistical evaluation by a randomization method, gave authentic alignment scores from 4 to 10 standard deviations above the means of the jumbled comparisons. The cumulative weight of the multiple alignment (Fig. 2) further bears out the significance of the overall relationship. That the polymerase portion of the viral reverse transcriptase system must encompass the amino-terminal portion of the 90-kDa fragment is established by the alignment shown in Fig. 3. The key region here involves a sector previously shown by Kamer and Argos (20) to be present in a number of nonretroviral polymerases; these consistently have two aspartic acid residues surrounded by...

▪ Abstract  Energy taxis is widespread in motile bacteria and in some species is the only known behavioral response. The bacteria monitor their cellular energy levels and respond to a decrease in energy by swimming to a microenvironment that reenergizes the cells. This is in contrast to classical Escherichia coli chemotaxis in which sensing of stimuli is independent of cellular metabolism. Energy taxis encompasses aerotaxis (taxis to oxygen), phototaxis, redox taxis, taxis to alternative electron acceptors, and chemotaxis to a carbon source. All of these responses share a common signal transduction pathway. An environmental stimulus, such as oxygen concentration or light intensity, modulates the flow of reducing equivalents through the electron transport system. A transducer senses the change in electron transport, or possibly a related parameter such as proton motive force, and initiates a signal that alters the direction of swimming. The Aer and Tsr proteins in E. coli are newly recognized transducers for energy taxis. Aer is homologous to E. coli chemoreceptors but unique in having a PAS domain and a flavin-adenine dinucleotide cofactor that is postulated to interact with a component of the electron transport system. PAS domains are energy-sensing modules that are found in proteins from archaea to humans. Tsr, the serine chemoreceptor, is an independent transducer for energy taxis, but its sensory mechanism is unknown. Energy taxis has a significant ecological role in vertical stratification of microorganisms in microbial mats and water columns. It plays a central role in the behavior of magnetotactic bacteria and also appears to be important in plant-microbe interactions.

ShaEP is a tool for rigid-body superimposition and similarity evaluation of ligand-sized molecules. Molecular overlay methods traditionally work on either substructures, molecular surfaces or interaction fields, or atom-centered Gaussian functions representing the molecular volume. While substructure searches are unlikely to reveal hits that are chemically different from the template structure, the other methods are capable of "scaffold hopping". Methods that match characteristic points in interaction fields can find alignments in situations where only some portions of the structures match but potentially miss good alignments if the used point sets are not detailed enough, which in turn increases the runtime of the used graph algorithms beyond practical limits. The faster, polynomially scaling volumetric methods consider the whole space to be equally important, which works well for molecules of equal size but partial matches might go undetected. ShaEP aims to capture the strengths of both field-based and volumetric approaches. It generates initial superimpositions using a matching algorithm on graphs that coarsely represent the electrostatic potential and local shape at points close to the molecular surfaces. The initial alignments are then optimized by maximization of the volume overlap of the molecules, computed using Gaussian functions. ShaEP overlays drug-sized molecules on a subsecond timescale, allowing for the screening of large virtual libraries. The program is available free of charge from www.abo.fi/fak/mnf/bkf/research/johnson/software.php.

BODIL is a molecular modeling environment geared to help the user to quickly identify key features of proteins critical to molecular recognition, especially (1) in drug discovery applications, and (2) to understand the structural basis for function. The program incorporates state-of-the-art graphics, sequence and structural alignment methods, among other capabilities needed in modern structure-function-drug target research. BODIL has a flexible design that allows on-the-fly incorporation of new modules, has intelligent memory management, and fast multi-view graphics. A beta version of BODIL and an accompanying tutorial are available at http://www.abo.fi/fak/mnf/bkf/research/johnson/bodil.html.