Contributions to microbiology最新文献

Antimicrobial peptides (AMPs) form a crucial part of human innate host defense, especially in neutrophil phagosomes and on epithelial surfaces. Bacteria have a variety of efficient resistance mechanisms to human AMPs, such as efflux pumps, secreted proteases, and alterations of the bacterial cell surface that are aimed to minimize attraction of the typically cationic AMPs. In addition, bacteria have specific sensors that activate AMP resistance mechanisms when AMPs are present. The prototypical Gram-negative PhoP/PhoQ and the Gram-positive Aps AMP-sensing systems were first described and investigated in Salmonella typhimurium and Staphylococcus epidermidis, respectively. Both include a classical bacterial two-component sensor/regulator system, but show many structural, mechanistic, and functional differences. The PhoP/PhoQ regulon controls a variety of genes not necessarily limited to AMP resistance mechanisms, but apparently aimed to combat innate host defense on a broad scale. In contrast, the staphylococcal Aps system predominantly upregulates AMP resistance mechanisms, namely the D-alanylation of teichoic acids, inclusion of lysyl-phosphati-dylglycerol in the cytoplasmic membrane, and expression of the putative VraFG AMP efflux pump. Notably, both systems are crucial for virulence and represent possible targets for antimicrobial therapy.

The PEP-dependent carbohydrate:phosphotransferase systems (PTSs) of enteric bacteria constitute a complex sensory system which involves as its central element a PEP-dependent His-protein kinase (Enzyme I). As a unit, the PTS comprises up to 20 different transporters per cell which correspond to its chemoreceptors for PTS carbohydrates, and several targeting subunits, which include in the low [G+C] Gram-positive bacteria an ancillary Ser/Thr-protein kinase. The PTS senses the presence of carbohydrates, in particular glucose, in the medium and the energy state of the cell, in the form of either the intracellular PEP-to-pyruvate ratio or the D-fructose-bisphosphate levels. This information is subsequently communicated to cellular targets, in particular those involved in the chemotactic response of the cell towards PTS carbohydrates, and in sensing glucose in the medium, using cAMP and several targeting subunits as intermediates. Peptide targeting subunits ensure the fast, transient, and yet accurate communication of the PTS with its more than hundred different targets, avoiding at the same time unwanted cross-talk. Many elements of this sensory system are simultaneously elements of specific and global regulatory networks. Thus, the PTS controls, besides the immediate (in the ms to s range) chemotactic responses, the activity of the various carbohydrate transporters and enzymes involved in carbon and energy metabolism through inducer exclusion, and in a delayed response (in the min to h range) the synthesis of these transporters and catabolic enzymes through catabolite repression. Indirect consequences of this program are phenomena related to cell surface rearrangements, which include flagella synthesis, as well as memory, adaptation, and learning effects. The analogy between the PTS and other prokaryotic systems, and more complex sensory systems from eukaryotic organisms which share elements with regulatory systems is obvious.

Recently, the list of ubiquitous bacterial secondary messengers which include cAMP and ppGpp has been extended by 3',5'-cyclic diguanylic acid (c-di-GMP). C-di-GMP metabolism is tuned by the tightly controlled activity of diguanylate cyclases and c-di-GMP-specific phosphodiesterases. As c-di-GMP-metabolizing enzymes are not only found frequently in bacterial genomes, but also are often numerous in individual genomes, the c-di-GMP metabolic network is highly complex whereby signaling specificity is adjusted on the level of expression, enzymatic activity, protein localization and, most likely, receptor affinity. The targets of c-di-GMP, which include protein and RNA receptors, are subsequently being unraveled. Besides the transition between sessility and motility, probably the most ancient regulatory control of bacterial behavior by c-di-GMP, many more phenotypes such as virulence are affected by c-di-GMP. However, the exact molecular mechanisms of c-di-GMP action remain to be discovered.

Magnetotactic bacteria orient and migrate along geomagnetic field lines. Magneto-aerotaxis increases the efficiency of respiring microaerophilic cells to efficiently find and maintain a position at a preferred microaerobic oxygen concentration. Magneto-aerotaxis could also facilitate access to regions of higher nutrient and electron acceptor concentration via periodic excursions above and below the preferred oxygen concentration level.

In the emerging field of synthetic biology, a central goal is to reliably engineer bacteria to respond to environmental signals according to a pre-determined genetic program. The sensor systems and genetic circuitry inside bacteria are the 'eyes' and 'brain' of a new class of biotechnological applications in which bacteria are used as living, self-replicating computers that can beneficially interact with the physical world. These engineered gene networks are constructed by extracting natural sensor systems and other genetic parts from multiple organisms and recombining them into novel configurations. This chapter is a how-to guide. It describes several strategies for engineering new bacterial sensor systems and synthetic gene networks that are capable of sensing a desired stimulus and generating interesting dynamical or pattern-forming responses. We also provide specification sheets describing many two-component and quorum-sensing systems, focusing on the information that one needs to know in order to use them for engineering applications.

Bacteria have developed several mechanisms which allow the preferred utilization of the most efficiently metabolizable carbohydrates when these organisms are exposed to a mixture of carbon sources. Interestingly, the same or similar mechanisms are used by some pathogens to control various steps of their infection process. The efficient metabolism of a carbon source might serve as signal for proper fitness. Alternatively, the presence of a specific carbon source might indicate to bacterial cells that they thrive in infection-related organs, tissues or cells and that specific virulence genes should be turned on or switched off. Frequently, virulence gene regulators are affected by changes in carbon source availability. For example, expression of the gene encoding the Streptococcus pyogenes virulence regulator Mga is controlled by the classical carbon catabolite repression (CCR) mechanism operative in Firmicutes. The activity of PrfA, the major virulence regulator in Listeria monocytogenes, seems to be controlled by the phosphorylation state of phosphotransferase system(PTS) components. In Vibrio cholerae synthesis of HapR, which regulates the expression of genes required for motility, is controlled via the Crp/cAMP CCR mechanism, whereas synthesis of Salmonella enterica HilE, which represses genes in a pathogenicity island, is regulated by the carbohydrate-responsive, PTS-controlled Mlc.

Cell-cell communication in bacteria, called quorum sensing, relies on production, release, and detection of signaling molecules, termed autoinducers. Communication enables populations of cells to synchronize gene expression and therefore behave as a group in a manner akin to cells in multicellular organisms. Most quorum-sensing systems allow communication within an individual species of bacteria. However, one autoinducer, called AI-2, is produced and recognized by many different bacterial species, indicating that some bacteria communicate across species boundaries. Current studies are aimed at discovering the role that AI-2 plays in gene regulation. Differential gene expression in response to AI-2 may cause bacterial behavioral changes, such as biofilm formation or transition to a pathogenic state. Interestingly, multiple mechanisms to detect AI-2 exist. These differences likely reflect variations in the role that AI-2 plays for different bacteria. Additionally, structural analyses of the AI-2 receptor in V. harveyi have provided insight into bacterial trans-membrane signal transduction. A further understanding of bacterial quorum-sensing processes may facilitate development of new technologies aimed at interfering with bacterial communication and virulence.

Global regulation of virulence gene expression via transcriptional regulators plays a central role in the ability of the bacterial pathogen Streptococcus pyogenes (the group A Streptococcus, GAS) to rapidly adapt during infection. The 'stand-alone' regulators Mga, RofA-like proteins (RALPs), and RopB/Rgg control important and diverse virulence regulons in response to growth-related signals and other environmental conditions in GAS. Stand-alone regulated genes encode factors important for colonization of tissues, immune evasion, persistence, dissemination, metabolism, and the response to stressors. Although conserved 'core' regulons have been established for each, recent studies have revealed significant inter-serotype and even intra-serotype variation in the regulatory patterns presented by the stand-alone regulators. This chapter will look at each stand-alone regulatory pathway in depth and discuss how these important global networks influence virulence as well as interact with each other to produce an integrated response during GAS infection.

During the course of an infection, a pathogenic bacterium has to sense the environment and adjust its gene expression appropriately. One such environmental cue is the difference in temperature inside and outside the host. RNA thermosensors are structures that can respond to differences in temperature by altering their conformation and thereby allowing/preventing binding of the ribosome to the translational start site. This chapter discusses different types of RNA thermosensors in general and RNA thermosensors known to control virulence gene expression in particular.