1 Center for Biological Sequence Analysis, BioCentrum-DTU, Building 208, The Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark
2 Molecular Microbiology and Genomics Consultants, Zotzenheim, Germany
3 Genomics & Strain Development, Chr. Hansen A/S, Hørsholm, Denmark
4 Microbial Adhesion Group, Center for Biomedical Microbiology, BioCentrum-DTU, Technical University of Denmark, Denmark
Correspondence
David W. Ussery
(dave{at}cbs.dtu.dk)
Genomes of the month
Seven new microbial genomes have been published since last month's Genome Update column was written. As usual, there is a heavy bias towards members of the Proteobacteria (which constitute about half of all the bacterial genomes sequenced so far; 107 out of 225). The list of new genomes includes five Proteobacteria. Two are members of the -Proteobacteria, the rickettsia Ehrlichia ruminantium, and Gluconobacter oxydans, which is of interest to the food industry. Three are
-Proteobacteria, the warfare germ Francisella tularensis, the plant pathogen Xanthomonas oryzae and finally Vibrio fischeri. The other genomes are of the probiotic Lactobacillus acidophilus and the archaeon Thermococcus kodakaraensis. A brief overview of these genomes is given below and a summary is presented in Table 1
.
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Gluconobacter oxydans belongs to the family Acetobacteraceae. These organisms have been used since ancient times in biotechnological processes like the production of vinegar and are still used for industrial applications which take advantage of their ability to incompletely oxidize a great variety of carbohydrates, alcohols and related compounds. The genome of Gluconobacter oxydans strain 621H (Prust et al., 2005) consists of one main 2·7 Mb circular chromosome and an additional five plasmids (pGOX1pGOX5, ranging from 163 to 2·6 kb), comprising 232 ORFs. The genome has an A+T content of 39 mol% and contains 2432 ORFs (1877 of these have an assigned function). The unique metabolism of Gluconobacter oxydans makes it an ideal model organism to study microbial processing of food.
Lactobacillus acidophilus is a representative member of the lactic acid bacteria, which are used in food and feed fermentations, such as dairy and silage. Lactic acid bacteria include probiotic strains, several of which have been sequenced. The 2·0 Mb genome of Lactobacillus acidophilus NCFM (Altermann et al., 2005) with 1864 predicted CDSs is relatively small and also has limited biosynthetic capabilities; most amino acids, cofactors and vitamins cannot be synthesized. On the other hand, the genome contains a relatively large fraction of genes involved in taking up various sugars and amino acids/peptides. This is consistent with its relatively nutrient-rich habitat of the gastrointestinal (GI) tract. The complex sugar fructooligosaccharide (FOS), shown to promote growth of probiotic species in the GI tract, can be metabolized by Lactobacillus acidophilus NCFM.
As with other sequenced bacteria, a large fraction of the predicted genes have no known function. Subsets of genes, for example those involved in adhesion, may be of particular interest because probiotic species must interact in various ways with the epithelial cells of the GI tract to exert their various beneficial effects. Future research will be needed to identify if there are fundamental differences between adhesion and colonization properties of benign organisms compared to pathogens.
Thermococcus kodakaraensis is a hyperthermophilic archaeon which can reduce elemental sulfur during growth and lives in high-temperature ecosystems. T. kodakaraensis contains a single, circular chromosome of 2·1 Mb in which 2306 genes have been found, covering 92 % of the genome, with a mean length of 833 bp. The genera of Thermococcus and Pyrococcus are closely related and both belong to the euryarchaeal order Thermococcales. A comparison of T. kodakaraensis to the Pyrococcus genomes of Pyrococcus horikoshii, Pyrococcus furiosus and Pyrococcus abyssi revealed 1204 shared proteins. The A+T content of T. kodakaraensis (48 mol%) is lower than the observed 5560 mol% for the three Pyrococcus genomes (Fukui et al., 2005
).
Vibrio fischeri is a species of bioluminescent bacteria that exists naturally in a free-living planktonic state, as a symbiont of certain luminescent fishes or in squid. Genome sequences of several Vibrio species that cause human diseases (Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus) have already been reported. The sequenced V. fischeri strain (ES114) is a representative of a non-pathogenic species (Ruby et al., 2005). Its genome is 4·3 Mb and is divided into two chromosomes and a 45·8 kb plasmid (pES100). V. fischeri strain ES114 has an A+T content of 62 mol% and contains 3802 predicted genes. Most rRNAs and tRNAs (11 and 108, respectively) were found in chromosome I, although chromosome II encodes a single rRNA operon and 11 tRNA genes. Despite V. fischeri ES114 being considered non-pathogenic, by comparing and analysing the sequence, interesting parallels with Vibrio cholerae and other pathogens were found.
Xanthomonas oryzae is a -proteobacterium belonging to the pathovar oryzae, responsible for bacterial blight (BB) of rice (Lee et al., 2005
). This phytopathogenic strain causes endemic disease in tropical Asian countries. The X. oryzae strain KACC10331 genome is 4·94 Mb long with an A+T content of 36 mol% and 4637 predicted genes. Comparisons with two non-pathogenic Xanthomonas strains sequenced previously revealed 245 species-specific genes in pathogenic Xanthomonas oryzae strain KACC10331. This strain contains twice as many transposable elements as the two other sequenced Xanthomonas strains. Although a large number of genes have already been characterized, more work is needed to understand the many aspects of virulence mechanisms of this important plant pathogen.
Method of the month prediction of protein secretion systems in bacterial genomes
For most bacteria it is essential to secrete particular proteins, and there are several methods available for predicting which proteins will be secreted and how. This genome update deals with characterization of secretion systems in bacterial genomes. Next month, the prediction of secreted proteins will be described, and the following month we will look at prediction of membrane proteins and combining all this to characterize where all the proteins in a given bacterial proteome are likely to be localized for Gram-negative bacteria, there are five possibilities: the cytosol (inside), embedded in the inner membrane, periplasm, outer membrane or secreted (outside).
This month the focus is on the different bacterial secretion systems (types IV). A database was constructed which includes information for Gram-positive and Gram-negative bacteria (J. D. Bendtsen, T. T. Binnewies, P. F. Hallin & D. W. Ussery, unpublished). We found most of the proteins for secretion systems type IV manually by screening the UniProt database, since this database contains only proteins with experimentally verified function. It is important to emphasize that currently we do not cover all available proteins for each individual secretion system. We are still developing and expanding the secretion system database and we will continue work to update it.
A more detailed analysis of the secretome (all proteins involved in secretion) will be given elsewhere (J. D. Bendtsen, T. T. Binnewies, P. F. Hallin & D. W. Ussery, unpublished) and will be only briefly sketched here. After collecting all homologues of the individual components of the five different secretion systems from UniProt, a Hidden Markov model (HMM) was built to specify conserved sequences for each individual secretion system. The individual HMMs were used to search all available proteomes in our bacterial database. The number of hits' for a defined cut-off value for each secretion system was counted and stored in the database. A preliminary version of the secretion type database is available on our supplementary web page (http://www.cbs.dtu.dk/services/GenomeAtlas/suppl/GenUp015/). Fig. 1 shows a difference in the distribution of the various secretion systems between Gram-positive and Gram-negative bacteria. For the Gram-positive organisms almost no type II or III secretion systems could be identified with the marked exception of Symbiobacterium thermophilum, which contains a type III secretion system (TTSS). Interestingly, according to Ueda et al. (2004)
the phylogeny derived from 16S rRNA suggests that this bacterium belongs to an unknown taxon in the Gram-positive bacterial cluster. In addition, a TTSS was predicted, perhaps assembled from Fli and FlhA/B proteins associated with flagellum assembly. These data are in agreement with the results from our secretion system database. This illustrates how genome sequences can raise, or answer, questions on taxonomic divisions of bacteria.
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Acknowledgements
This work was supported by a grant from the Danish Center for Scientific Computing.
REFERENCES
Altermann, E., Russell, W. M., Azcarate-Peril, M. A. & 11 other authors (2005). Complete genome sequence of the probiotic lactic acid bacterium Lactobacillus acidophilus NCFM. Proc Natl Acad Sci U S A (in press). doi:10.1073/pnas.0409188102
Collins, N. E., Liebenberg, J., De Villiers, E. P. & 19 other authors (2005). Genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number. Proc Natl Acad Sci U S A 102, 838843.
Fukui, T., Atomi, H., Kanai, T. Matsumi R., Fujiwara, S. & Imanaka, T. (2005). Complete genome sequence of the hyperthermophilic archaeon Thermococcus kodakaraensis Kod1 and comparison with Pyrococcus genomes. Genome Res 15, 352363.
Larsson, P., Oyston, P. C., Chain, P. & 24 other authors (2005). The complete genome sequence of Francisella tularensis, the causative agent of tularemia. Nat Genet 37, 153159.[CrossRef][Medline]
Lee, B. M., Park, Y. J., Park, D. S. & 16 other authors (2005). The genome sequence of Xanthomonas oryzae pathovar oryzae KACC10331, the bacterial blight pathogen of rice. Nucleic Acids Res 33, 577586.
Prust, C., Hoffmeister, M., Liesegang, H., Wiezer, A., Fricke, W. F., Ehrenreich, A., Gottschalk, G. & Deppenmeier, U. (2005). Complete genome sequence of the acetic acid bacterium Gluconobacter oxydans. Nat Biotechnol 23, 195200.[CrossRef][Medline]
Ruby, E. G., Urbanowski, M., Campbell, J. & 13 other authors (2005). Genome sequence of Vibrio fischeri: a symbiotic bacterium with pathogenic congeners. Proc Natl Acad Sci U S A 102, 30043009.
Willenbrock, H., Binnewies, T. T., Hallin, P. F. & Ussery, D. W. (2005). Genome Update: 2D clustering of bacterial genomes. Microbiology 151, 333336.[CrossRef][Medline]
Ueda, K., Yamashita, A., Ishikawa, J., Shimada, M., Watsuji, T. O., Morimura, K., Ikeda, H., Hattori, M. & Beppu, T. (2004). Genome sequence of Symbiobacterium thermophilum, an uncultivable bacterium that depends on microbial commensalism. Nucleic Acids Res 32, 49374944.
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