Initially, most eukaryotic sequence projects were devoted to typical animals, fungi and plants. Now more and more effort is put into sequencing protist genomes. Protists are an artificial assemblage containing mostly unicellular eukaryotes from different phylogenetic lineages, and are much more diversified and widespread than higher Eukaryota. Sequenced protist genomes are essential for reconstructing the Tree of Life and understanding significant events in eukaryotic evolution and diversification. Many protists are parasites and pathogens with medical and economic significance, while others play important ecological roles as primary producers and crucial links in food webs. A number of protists also serve as model organisms in various biological fields and are becoming important in biotechnology. Thirty-seven protist genomes were published by the beginning of 2010 and 217 projects are ongoing. Knowledge coming from these projects will be helpful in developing more efficient protection from pathogenic protists and their effective elimination. Sequenced genomes of ecologically important protists could help to understand key environmental phenomena and even to control them. Newly sequenced protist genomes encode previously unknown enzymes and metabolic pathways, which will be useful in further development of biotechnology.
Initiation of bacterial chromosome replication is mediated by a single initiator protein ? DnaA which interacts specifically with multiple DnaA boxes located within the origin of replication oriC. We have applied in silico methods: DNA asymmetry, DnaA box distribution and dnaA gene location to identify the putative replication origins in bacterial chromosomes. The three methods identify the same region as a putative origin in more than half of the analyzed chromosomes. The most universal method of putative oriC identification in bacterial chromosomes is DNA asymmetry, although in some cases it is necessary to apply all three methods. Interestingly, most bacterial chromosomes exhibit an overrepresentation of DnaA boxes; they contain at least one cluster of DnaA boxes in the vicinity of the oriC region that is probably involved in controlling replication initiation.
We have analysed the coding capacity of ORFs longer than 100 codons found in the yeast genome. Comparing the parameters describing the DNA asymmetry in the set of known genes and the set of all ORFs>100 codons we have found that there are about 4700 coding ORFs in the yeast genome. Since for more than 2300 ORFs recognisable functions have been already found and for about 2000 ORFs homology to known genes has been identified - only about 400 ORFs can be considered as orphans - ORFs without any known function or homology. This finding means that there is no mystery of orphans - a paradox showing that the fraction of orphans has been growing with the growing number of genes with known functions in the yeast genome.
Genomics is a new field of biology. Its fast development is caused mainly by quick progress in large-scale genome sequencing and in computer technology. In spite of a huge number of sequenced microbial genomes available in databases, their taxonomical diversity is biased and reflects the interests of researches and facility of microorganisms' isolation and culture in laboratory conditions. More than 80% of genome sequencing projects are focused on the members of Proteobacteria, Firmicutes and Actinobacteria. Environmental genome shotgun sequencing reveals that microbial diversity is much greater than we expected. Particular levels of genomic analysis, the problems and subjects of genomics are specified and described here.
Availability of fully sequenced genomes contributes to the development of new science named phylogenomics which opens new possibilities of phylogenetic analyses and study of genome evolution based on the whole information coded in genomic DNA. The advantages and disadvantages of the new methods are described. Despite many phenomena such as lineage-specific gene loss, gene duplication and horizontal gene transfer disturbing phylogenetic analyses, the new methods are able to extract some phylogenetic signals in the analysed genomes and construct reliable phylogenetic trees. The genome-based studies support not only the three-domain concept of Tree of Life but they identify previously undetected major clades of prokaryotes as well.
Today, we have very powerful and effective machines and methods to sequence and analyze DNA sequences. Almost every week, new genomes are added to sequence databases. However, those data are useless without additional annotations. Genes need to be found and their functions defined. Experimental work is too slow to analyze each sequence of a potential gene but computational methods facilitate such analyses. Here, we review the methodology, potential problems and constraints in genes finding and their annotation. We describe some new approaches including comparative genomics.
In bacterial chromosomes, strong bias in nucleotide composition has been observed between differently replicated DNA strands (leading and lagging ones), and also in many species between the regions proximal and distal to the origin of replication (ori). This bias is also reflected in composition and distribution of genes along the chromosome. Several phenomena connected with the replication of the chromosome are responsible for such polarization, especially mutational pressure, repair mechanisms and recombinations, and also selection pressure. All these phenomena are not indifferent for gene evolution and their rearrangements which are strictly connected with the organization of bacterial chromosome.
We have analysed the causes of asymmetry in nucleotide composition of DNA complementary strands of prokaryotic chromosomes. Analysing DNA walks we have separated the effect of replication-associated processes from the effect introduced by transcription and coding functions. The asymmetry introduced by replication switches its polarity at the origin and at the terminus of replication, which is observed in both noncoding and coding sequences and varies with respect to positions in codons. Coding functions introduce very strong trends into protein coding ORFs, which are specific for each nucleotide position in the codon. Using detrended DNA walks we have eliminated the effect of coding density and we were able to distinguish between mutational pressure associated with replication and compositional bias for genes proximal and distal to the origin of replication.
One of the common features of bacterial genomes is a strong compositional asymmetry between differently replicating DNA strands (leading and lagging). The main cause of the observed bias is the mutational pressure associated with replication. This suggests that genes translocated between differently replicating DNA strands are subjected to a higher mutational pressure, which may influence their composition and divergence rate. Analyses of groups of completely sequenced bacterial genomes have revealed that the highest divergence rate is observed for the DNA sequences that in closely related genomes are located on different DNA strands in respect to their role in replication. Paradoxically, for this group of sequences the absolute values of divergence rate are higher for closely related species than for more diverged ones. Since this effect concerns only the specific group of orthologs, there must be a specific mechanism introducing bias into the structure of chromosome by enriching the set of homologs in trans position in newly diverged species in relatively highly diverged sequences. These highly diverged sequences may be of varied nature: (1) paralogs or other fast-evolving genes under weak selection; or (2) pseudogenes that will probably be eliminated from the genome during further evolution; or (3) genes whose history after divergence is longer than the history of the genomes in which they are found. The use of these highly diverged sequences for phylogenetic analyses may influence the topology and branch length of phylogenetic trees. The changing mutational pressure may contribute to arising of genes with new functions as well.
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