RESUMO
We describe here the complete genome sequence (1,111,523 base pairs) of the obligate intracellular parasite Rickettsia prowazekii, the causative agent of epidemic typhus. This genome contains 834 protein-coding genes. The functional profiles of these genes show similarities to those of mitochondrial genes: no genes required for anaerobic glycolysis are found in either R. prowazekii or mitochondrial genomes, but a complete set of genes encoding components of the tricarboxylic acid cycle and the respiratory-chain complex is found in R. prowazekii. In effect, ATP production in Rickettsia is the same as that in mitochondria. Many genes involved in the biosynthesis and regulation of biosynthesis of amino acids and nucleosides in free-living bacteria are absent from R. prowazekii and mitochondria. Such genes seem to have been replaced by homologues in the nuclear (host) genome. The R. prowazekii genome contains the highest proportion of non-coding DNA (24%) detected so far in a microbial genome. Such non-coding sequences may be degraded remnants of 'neutralized' genes that await elimination from the genome. Phylogenetic analyses indicate that R. prowazekii is more closely related to mitochondria than is any other microbe studied so far.
Assuntos
Evolução Molecular , Genoma Bacteriano , Mitocôndrias/genética , Rickettsia prowazekii/genética , Replicação do DNA , DNA Bacteriano , DNA Mitocondrial , Proteínas de Membrana/genética , Biossíntese de Proteínas , Recombinação Genética , Sequências Reguladoras de Ácido Nucleico , Sequências Repetitivas de Ácido Nucleico , Origem de Replicação , Rickettsia prowazekii/patogenicidade , Transcrição Gênica , Virulência/genéticaRESUMO
We describe here the first general survey of the genomic content and the coding capacity of the 1.1 Mb genome of Rickettsia prowazekii based on an analysis of a total of 200 kb of unique sequence data collected in a random manner. Based on nucleotide distribution profiles, we estimate that the R. prowazekii genome may have a coding density of 60%-70% and that it may contain a total of circa 800 genes. Here, we have tentatively identified and classified 173 of these genes. Our analysis suggests that the R. prowazekii genome is a highly derived, reduced genome that has lost many genes involved in amino acid biosynthetic pathways and regulatory functions. Furthermore, the R. prowazekii genome seems to lack glycolytic genes, but it does contain genes encoding components of the tricarboxylic acid cycle as well as of the electron transport system. We have also encountered a family of homologous genes coding for ATP/ADP translocases, as observed in several mitochondrial genomes. We relate these findings to previous phylogenetic studies that suggest that Rickettsia and mitochondria share a common ancestor.
Assuntos
Genoma Bacteriano , Rickettsia prowazekii/genética , Sequência de Bases , Ciclo do Ácido Cítrico/genética , DNA Mitocondrial/genética , Transporte de Elétrons/genética , Genes Bacterianos/genética , Translocases Mitocondriais de ADP e ATP/genética , Dados de Sequência Molecular , Filogenia , Análise de SequênciaRESUMO
We have previously shown that in Escherichia coli the accumulation of five leucine and three methionine tRNA species is regulated so that those tRNA species that translate major codons increase while those that translate minor codons decrease as the growth rate increases. Here, we have analyzed the growth-rate-dependence of another 12 tRNA species. We find that the level of three tRNA species cognate to the major glycine, proline and arginine codons, respectively, increase with increasing growth rates. Conversely, four tRNAs that are cognate to minor codons within the same amino acid families decrease with increasing growth rates. In addition, the glutamyl as well as the phenylalanyl isoacceptor species are accumulated in proportion to the content of these two amino acids in the proteins produced at different growth rates. In summary, the patterns of the growth-rate-dependence for the accumulation of these 17 tRNA species support the interpretation that the major codon preference is an arrangement to maximize the growth rates of bacteria in rich media by optimizing the kinetic efficiency of translation. In contrast, we find that three minor tRNA species cognate to two rare arginine codons and one minor glycine codon, respectively, increase with increasing growth rate. Such findings suggest that there are additional constraints on the accumulation of these tRNA species that may be distinct from those required to optimize the kinetic efficiency of translation.
Assuntos
Escherichia coli/crescimento & desenvolvimento , RNA de Transferência/metabolismo , Sequência de Bases , Northern Blotting , Códon , Escherichia coli/genética , Escherichia coli/metabolismo , Cinética , Dados de Sequência Molecular , Sondas de Oligonucleotídeos , Biossíntese de Proteínas , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , RNA de Transferência/genéticaRESUMO
We have used an affinity electrophoresis assay which when combined with Northern hybridization techniques permits us to estimate the degree of thiolation of individual tRNA species in Escherichia coli. We observe that the levels of 4-thio 2'(3')-uridine (4-thioU) in many but not all tRNAs varies dramatically at different bacterial growth rates: Five tRNAs are completely thiolated at all growth rates, while another eight tRNAs are incompletely thiolated and the fraction of the unthiolated form of these tRNA species increases as the growth rates increase. Transfer RNA(2Glu) contains 4-thioU as well as (methylamino)methyl-2-thio uridine (mnm(5)2-thioU). The level of mnm(5)2-thioU of tRNA(2Glu) is invariant with growth rate. Surprisingly, none of the thirteen tRNA species that we have studied is completely unmodified in all growth media. In particular, at the slowest growth rates every tRNA class that we have studied contains a form that has 4-thioU residues.