GTPases are conserved molecular switches, built according to a common structural design. Rapidly accruing knowledge of individual GTPases--crystal structures, biochemical properties, or results of molecular genetic experiments--support and generate hypotheses relating structure to function in other members of the diverse family of GTPases.

The GTPase superfamily: conserved structure and molecular mechanism

Comparative sequence analysis of small subunit rRNA is currently one of the most important methods for the elucidation of bacterial phylogeny as well as bacterial identification. Phylogenetic investigations targeting alternative phylogenetic markers such as large subunit rRNA, elongation factors, and ATPases have shown that 16S rRNA-based trees reflect the history of the corresponding organisms globally. However, in comparison with three to four billion years of evolution the phylogenetic information content of these markers is limited. Consequently, the limited resolution power of the marker molecules allows only a spot check of the evolutionary history of microorganisms. This is often indicated by locally different topologies of trees based on different markers, data sets or the application of different treeing approaches. Sequence peculiarities as well as methods and parameters for data analysis were studied with respect to their effects on the results of phylogenetic investigations. It is shown that only careful data analysis starting with a proper alignment, followed by the analysis of positional variability, rates and character of change, testing various data selections, applying alternative treeing methods and, finally, performing confidence tests, allows reasonable utilization of the limited phylogenetic information.

Bacterial phylogeny based on comparative sequence analysis.

Molecular phylogeny increasingly supports the understanding of organismal relationships and provides the basis for the classification of microorganisms according to their natural affiliations. Comparative sequence analysis of ribosomal RNAs or the corresponding genes currently is the most widely used approach for the reconstruction of microbial phylogeny. The highly and less conserved primary and higher order structure elements of rRNAs document the history of microbial evolution and are informative for definite phylogenetic levels. An optimal alignment of the primary structures and a careful data selection are prerequisites for reliable phylogenetic conclusions. rRNA based phylogenetic trees can be reconstructed and the significance of their topologies evaluated by applying distance, maximum parsimony and maximum likelihood methods of phylogeny inference in comparison, and by fortuitous or directed resampling of the data set. Phylogenetic trees based on almost equivalent data sets of bacterial 23S and 16S rRNAs are in good agreement and their overall topologies are supported by alternative phylogenetic markers such as elongation factors and ATPase subunits. Besides their phylogenetic information content, the differently conserved primary structure regions of rRNAs provide target sites for specific hybridization probes which have been proven to be powerful tools for the identification of microbes on the basis of their phylogenetic relationships.

Bacterial phylogeny based on 16S and 23S rRNA sequence analysis

Introduction to the Proteobacteria

α-l-Arabinofuranosidases: biochemistry, molecular biology and application in biotechnology

Comparative Sequence Analysis of 23S rRNA from Proteobacteria

The gene coding for the elongation factor Tu (EF-Tu) of Thermatoga maritima was cloned and sequenced. The predicted amino acid sequence was compared with those of other eubacteria, an archaebacterium and two eukaryotes as well. The similarity values and the distance matrix tree show that Thermotoga is more closely related to the eubacteria than to the representatives of the other urkingdoms. Thermotoga maritima represents the deepest branching within the tree of EF-Tu sequences from all eubacteria studied so far.

Nucleotide sequence of the gene coding for the elongation factor Tu from the extremely thermophilic eubacterium Thermotoga maritima

The nucleotide sequence of the α-l-arabinofuranosidase gene arfB from Clostridium stercorarium was determined. The deduced protein has a molecular mass of 56.2 kDa with an amino terminus identical to the N-terminal sequence of the purified mature enzyme from C. stercorarium. Its sequence is homologous to arabinofuranosidases of glycosyl hydrolase family 51. Sequence alignment and cluster analysis reveal three new members of glycosyl hydrolase family 51, allowing for the definition of highly conserved regions. Two of these regions are remarkably similar to the most conserved regions within several other families of glycosyl hydrolases, which have in common a (β/α)8-barrel as the core super-secondary structure, and allow to predict the acid/base catalyst and the nucleophile of the active site.

Nucleotide sequence of arfB of Clostridium stercorarium, and prediction of catalytic residues of α-L-arabinofuranosidases based on local similarity with several families of glycosyl hydrolases

Marianne Bachleitner

Papers

Comparative Sequence Analysis of 23S rRNA from Proteobacteria

Nucleotide sequence of the gene coding for the elongation factor Tu from the extremely thermophilic eubacterium Thermotoga maritima

Nucleotide sequence of arfB of Clostridium stercorarium, and prediction of catalytic residues of α-L-arabinofuranosidases based on local similarity with several families of glycosyl hydrolases