RESUMO
ßlactam resistance in clinical isolates of Streptococcus pneumoniae arises by only one route, the reduction of the affinity of the penicillin-binding proteins (PBPs) for ßlactams. The pneumococcus possesses five high molecular weight PBPs (PBP1A, 1B, 2A, 2B, and 2X) which are involved in the final crosslinking stages of peptidoglycan synthesis in the bacterial cell wall. ßlactam antibiotics are structural analogs of the natural cell wall peptide substrates of the PBPs. The antibiotic binds to the active site within the transpeptidase domain of these PBPs, forming an acyl-enzyme complex which is far more stable than the transient enzyme-substrate complex that normally occurs. In this way, the ßlactams block the crosslinking in what is essentially an irreversible manner. The result is a cessation in cell growth and, depending on the PBP inhibited, lysis.
RESUMO
The com operon of naturally transformable streptococcal species contains three genes, comC, comD, and comE, involved in the regulation of competence. The comC gene encodes a competence-stimulating peptide (CSP) thought to induce competence in the bacterial population at a critical extracellular concentration. The comD and comE genes are believed to encode the transmembrane histidine kinase and response regulator proteins, respectively, of a two-component regulator, with the comD-encoded protein being a receptor for CSP. Here we report on the genetic variability of comC and comD within Streptococcus pneumoniae isolates. Comparative analysis of sequence variations of comC and comD shows that, despite evidence for horizontal gene transfer at this locus and the lack of transformability of many S. pneumoniae strains in the laboratory, there is a clear correlation between the presence of a particular comC allele and the cognate comD allele. These findings effectively rule out the possibility that the presence of noncognate comC and comD alleles may be responsible for the inability to induce competence in many isolates and indicate the importance of a functional com pathway in these isolates. In addition, we describe a number of novel CSPs from disease-associated strains of S. mitis and S. oralis. The CSPs from these isolates are much more closely related to those from S. pneumoniae than to most CSPs previously reported from S. mitis and S. oralis, suggesting that these particular organisms may be a potential source of DNA in recombination events generating the mosaic structures commonly reported in genes of S. pneumoniae that are under strong selective pressure.
Assuntos
Proteínas de Bactérias/genética , Variação Genética/genética , Complexos Multienzimáticos , Proteínas Quinases/genética , Proteínas/genética , Streptococcus pneumoniae/genética , Transformação Bacteriana , Alelos , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Sequência de Bases , Sítios de Ligação , Transferência Genética Horizontal , Genes Bacterianos/genética , Histidina Quinase , Dados de Sequência Molecular , Óperon/genética , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo Genético/genética , Proteínas Quinases/química , Proteínas Quinases/metabolismo , Alinhamento de Sequência , Streptococcus/genética , Streptococcus/patogenicidade , Streptococcus pneumoniae/enzimologiaRESUMO
In enteric bacteria three discrete families of type I restriction and modification systems (IA, IB and ID) are encoded by alleles of the serB-linked hsd locus. Probes specific for each of the three families were used to monitor the distribution of related systems in 37 of the 72 wild-type Escherichia coli strains comprising the ECOR collection. All 25 members of group A in this collection were screened; 12 were probe-positive, nine have hsd genes in the IA family, two in the IB and one in the ID. Twelve strains, representing all groups other than A, were screened; five were probe-positive, one has hsd genes in the IA family, one in the IB and three in the ID. The type ID genes are the first representatives of this family in E. coli, the probe-negative strains could have alternative families of hsd genes. The type IA and IB systems added at least five new specificities to the five already identified in natural isolates of E. coli. The distribution of alleles is inconsistent with the dendrogram of the bacterial strains derived from other criteria. This discrepancy and the dissimilar coding sequences of allelic hsd genes both imply lateral transfer of hsd genes.
Assuntos
Alelos , Desoxirribonucleases de Sítio Específico do Tipo I/genética , Escherichia coli/genética , Genes Bacterianos , Família Multigênica , DNA Metiltransferases Sítio Específica (Adenina-Específica)/genética , Sequência de Bases , Clonagem Molecular , DNA Bacteriano/genética , Escherichia coli/classificação , Escherichia coli/enzimologia , Escherichia coli/isolamento & purificação , Evolução Molecular , Variação Genética , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Seleção Genética , Especificidade da Espécie , Especificidade por Substrato , Transformação BacterianaRESUMO
Mosaic penicillin-binding proteins (PBP) 1A, 2X and 2B genes were cloned from four clinical isolates of Streptococcus pneumoniae with levels of susceptibility to penicillin ranging from 1.5 to 16 micrograms benzylpenicillin ml-1. In each instance it was possible to transform either the penicillin-sensitive laboratory strain R6 or a sensitive clinical isolate 110K/70 to the full level of penicillin resistance with these three penicillin-binding proteins alone. Until now it has not been possible to clearly determine whether alterations to PBP1A, 2X and 2B alone were sufficient to attain high level penicillin resistance.
