The bacterial cytoskeleton

Bacteria contain a complex cytoskeleton that is more diverse than previously thought. Recent research provides insight into how bacterial actins, tubulins, and ParA proteins participate in a variety of cellular processes.     

The bacterial cytoskeleton.

In recent years it has been shown that bacteria contain a number of cytoskeletal structures. The bacterial cytoplasmic elements include homologs of the three major types of eukaryotic cytoskeletal proteins (actin, tubulin, and intermediate filament proteins) and a fourth group, the MinD-ParA group, that appears to be unique to bacteria. The cytoskeletal structures play important roles in cell division, cell polarity, cell shape regulation, plasmid partition, and other functions. The proteins self-assemble into filamentous structures in vitro and form intracellular ordered structures in vivo. In addition, there are a number of filamentous bacterial elements that may turn out to be cytoskeletal in nature. This review attempts to summarize and integrate the in vivo and in vitro aspects of these systems and to evaluate the probable future directions of this active research field.     

Biosensor technology: technology push versus market pull

Biosensor technology is based on a specific biological recognition element in combination with a transducer for signal processing. Since its inception, biosensors have been expected to play a significant analytical role in medicine, agriculture, food safety, homeland security, environmental and industrial monitoring. However, the commercialization of biosensor technology has significantly lagged behind the research output as reflected by a plethora of publications and patenting activities. The rationale behind the slow and limited technology transfer could be attributed to cost considerations and some key technical barriers. Analytical chemistry has changed considerably, driven by automation, miniaturization, and system integration with high throughput for multiple tasks. Such requirements pose a great challenge in biosensor technology which is often designed to detect one single or a few target analytes. Successful biosensors must be versatile to support interchangeable biorecognition elements, and in addition miniaturization must be feasible to allow automation for parallel sensing with ease of operation at a competitive cost. A significant upfront investment in research and development is a prerequisite in the commercialization of biosensors. The progress in such endeavors is incremental with limited success, thus, the market entry for a new venture is very difficult unless a niche product can be developed with a considerable market volume.     

Axon guidance: push and pull with ephrins and GDNF

The pathfinding of motor axons is an important model system for understanding binary axon guidance decisions. Recent work has shown that GDNF attracts motor neuron growth cones, and interacts synergistically with ephrinAs on growth cone directionality.     

The push and pull of climate change causes heterogeneous shifts in avian elevational ranges

Projected effects of climate change on animal distributions primarily focus on consequences of temperature and largely ignore impacts of altered precipitation. While much evidence supports temperature‐driven range shifts, there is substantial heterogeneity in species' responses that remains poorly understood. We resampled breeding ranges of birds across three elevational transects in the Sierra Nevada Mountains, USA, that were extensively surveyed in the early 20th century. Presence–absence comparisons were made at 77 sites and occupancy models were used to separate significant range shifts from artifacts of false absences. Over the past century, rising temperature pushed species upslope while increased precipitation pulled them downslope, resulting in range shifts that were heterogeneous within species and among regions. While 84% of species shifted their elevational distribution, only 51% of upper or lower range boundary shifts were upslope. By comparison, 82% of range shifts were in a direction predicted by changes in either temperature or precipitation. Species were significantly more likely to shift elevational ranges than their ecological counterparts if they had small clutch sizes, defended all‐purpose territories, and were year‐round residents, results that were in opposition to a priori predictions from dispersal‐related hypotheses. Our results illustrate the complex interplay between species‐specific and region‐specific factors that structure patterns of breeding range change over long time periods. Future projections of increasing temperature and highly variable precipitation regimes create a strong potential for heterogeneous responses by species at range margins.     

Diversification and specialization of the bacterial cytoskeleton

The past decade has witnessed the identification and characterization of bacterial homologs of the three major eukaryotic cytoskeletal families: actin, tubulin and intermediate filaments. These proteins play essential roles in organizing bacterial subcellular environments. Recently, the ParA/MinD superfamily has emerged as a new bacterial cytoskeletal class, and imaging studies hint at the existence of even more, as yet unidentified, cytoskeletal systems. Much as the cytoskeleton is used for different purposes in different eukaryotic cells, the specific identities, functions and regulatory mechanisms of cytoskeletal proteins can vary between different bacterial species. In addition, extensive cross-talk between bacterial cytoskeletal systems may represent an important mode of cytoskeletal regulation. These themes of diversity, species-specificity and crosstalk are emerging as central properties of cytoskeletal biology.     

Increasing complexity of the bacterial cytoskeleton

Bacteria contain cytoskeletal elements involved in major cellular processes including DNA segregation and cell morphogenesis and division. Distant bacterial homologues of tubulin (FtsZ) and actin (MreB and ParM) not only resemble their eukaryotic counterparts structurally but also show similar functional characteristics, assembling into filamentous structures in a nucleotide-dependent fashion. Recent advances in fluorescence microscopic imaging have revealed that FtsZ and MreB form highly dynamic helical structures that encircle the cells along the inside of the cell membrane. With the discovery of crescentin, a cell-shape-determining protein that resembles eukaryotic intermediate filament proteins, the third major cytoskeletal element has now been identified in bacteria as well.     

A dynamic bacterial cytoskeleton

Actin and tubulin are the major components of the cytoskeleton that pervades the cytoplasm of all eukaryotic cells. These proteins were traditionally thought not to be present in prokaryotes, but structural and functional homologues of tubulin (FtsZ) and actin (MreB) are now known to be present virtually throughout the eubacteria and in some archae. FtsZ protein is a key player in cell division of bacteria and some eukaryotic organelles. MreB proteins are involved in the regulation of cell shape and the segregation of some bacterial plasmids, and might have a range of other functions. Recent data demonstrate that the bacterial proteins are, like their eukaryotic counterparts, highly dynamic. Here, we review the general properties and functions of actin and tubulin homologues in bacteria, their dynamic behaviour and the implications for understanding cell division and morphogenesis in bacteria.     

External push and internal pull forces recruit curvature-sensing N-BAR domain proteins to the plasma membrane

Many of the more than 20 mammalian proteins with N-BAR domains control cell architecture and endocytosis by associating with curved sections of the plasma membrane. It is not well understood whether N-BAR proteins are recruited directly by processes that mechanically curve the plasma membrane or indirectly by plasma-membrane-associated adaptor proteins that recruit proteins with N-BAR domains that then induce membrane curvature. Here, we show that externally induced inward deformation of the plasma membrane by cone-shaped nanostructures (nanocones) and internally induced inward deformation by contracting actin cables both trigger recruitment of isolated N-BAR domains to the curved plasma membrane. Markedly, live-cell imaging in adherent cells showed selective recruitment of full-length N-BAR proteins and isolated N-BAR domains to plasma membrane sub-regions above nanocone stripes. Electron microscopy confirmed that N-BAR domains are recruited to local membrane sites curved by nanocones. We further showed that N-BAR domains are periodically recruited to curved plasma membrane sites during local lamellipodia retraction in the front of migrating cells. Recruitment required myosin-II-generated force applied to plasma-membrane-connected actin cables. Together, our results show that N-BAR domains can be directly recruited to the plasma membrane by external push or internal pull forces that locally curve the plasma membrane.     

The spectrin cytoskeleton is crucial for adherent and invasive bacterial pathogenesis

Various enteric bacterial pathogens target the host cell cytoskeletal machinery as a crucial event in their pathogenesis. Despite thorough studies detailing strategies microbes use to exploit these components of the host cell, the role of the spectrin-based cytoskeleton has been largely overlooked. Here we show that the spectrin cytoskeleton is a host system that is hijacked by adherent (Entropathogenic Escherichia coli [EPEC]), invasive triggering (Salmonella enterica serovar Typhimurium [S. Typhimurium]) and invasive zippering (Listeria monocytogenes) bacteria. We demonstrate that spectrin cytoskeletal proteins are recruited to EPEC pedestals, S. Typhimurium membrane ruffles and Salmonella containing vacuoles (SCVs), as well as sites of invasion and comet tail initiation by L. monocytogenes. Spectrin was often seen co-localizing with actin filaments at the cell periphery, however a disconnect between the actin and spectrin cytoskeletons was also observed. During infections with S. Typhimurium ΔsipA, actin-rich membrane ruffles at characteristic sites of bacterial invasion often occurred in the absence of spectrin cytoskeletal proteins. Additionally, early in the formation of L. monocytogenes comet tails, spectrin cytoskeletal elements were recruited to the surface of the internalized bacteria independent of actin filaments. Further studies revealed the presence of the spectrin cytoskeleton during SCV and Listeria comet tail formation, highlighting novel cytoplasmic roles for the spectrin cytoskeleton. SiRNA targeted against spectrin and the spectrin-associated proteins severely diminished EPEC pedestal formation as well as S. Typhimurium and L. monocytogenes invasion. Ultimately, these findings identify the spectrin cytoskeleton as a ubiquitous target of enteric bacterial pathogens and indicate that this cytoskeletal system is critical for these infections to progress.     

Structural insights into bacterial modulation of the host cytoskeleton

Many bacterial pathogens manipulate the host cell cytoskeleton during infection. Such cytoskeletal modulation can occur at several points of contact between the pathogen and the host, and involves extracellular receptors, intracellular signal transduction and cytoskeletal proteins themselves. The field of bacterial pathogenesis has progressed dramatically over the past decade, such that structural knowledge is both timely and essential for a full appreciation of the biology at the pathogen-host interface. Several recent examples involving bacterial proteins that target actin, Rho family GTPases and extracellular receptors have contributed to a structural understanding of eukaryotic cytoskeletal modulation by pathogens.     

拒避-诱杀组合技术对橘小实蝇产卵的影响

本文研究了以矿物油(mineral spray oil)乳剂为保护剂、"猎蝇"蛋白质毒饵为诱杀剂组成的拒避-诱杀组合技术对橘小实蝇Bactrocera dorsalis (Hendel)产卵的控制作用。结果显示:橘小实蝇在采用拒避-诱杀组合技术保护的供试香蕉上平均产卵孔数为0.000±0.000个,果实完好率达到100%,而单独使用"猎蝇"蛋白质毒饵的平均产卵孔数为27.000±1.732个,果实完好率仅为5.55%,表明该组合技术对橘小实蝇产卵的控制效果显著。在100倍～300倍浓度范围内,矿物油浓度越高,拒避-诱杀组合对橘小实蝇产卵的控制效果越好;随着虫口密度的增加,毒饵使用量和施用面积也须相应增加,才能取得更好的防治效果。     

Uptake of cystic fibrosis testing in primary care: supply push or demand pull?

OBJECTIVE--To determine the acceptability and feasibility of screening for carriers of cystic fibrosis in a primary care setting. DESIGN--Follow up study over 15 months of patients offered carrier testing by mouthwash. SETTING--A general practice in inner London. SUBJECTS--5529 patients aged 18-45 invited by various methods and combinations of methods (letter, booklet, personal approach) for testing. MAIN OUTCOME MEASURES--Uptake of screening, anxiety, and knowledge of test. RESULTS--957 (17%) invitees were screened over the 15 months. 28 carriers and no carrier couples were detected. Uptake rates were 12% (59/502 patients) among patients invited by letter and tested by appointment; 9% (47/496) among patients invited by letter, with leaflet, and tested by appointment; 4% (128/2953) among patients invited by letter six weeks before the end of the study and tested by appointment; 17% (81/471) among patients offered passive opportunistic testing; 70% (453/649) among patients offered active opportunistic testing; and 25% (22/88) among patients offered active opportunistic testing by appointment. A short term rise in anxiety among those given a positive test result had dissipated by three months. At three months about one fifth and one third of those given positive and negative results respectively did not understand their results correctly. CONCLUSION--These results suggest that the strongest variable in determining uptake of screening is the active approach by a health professional offering immediate testing. It remains to be resolved whether the high uptake rates achieved by active recruitment indicate a supply push for this new test rather than a demand from the population.     

The bacterial cytoskeleton: an intermediate filament-like function in cell shape

Various cell shapes are encountered in the prokaryotic world, but how they are achieved is poorly understood. Intermediate filaments (IFs) of the eukaryotic cytoskeleton play an important role in cell shape in higher organisms. No such filaments have been found in prokaryotes. Here, we describe a bacterial equivalent to IF proteins, named crescentin, whose cytoskeletal function is required for the vibrioid and helical shapes of Caulobacter crescentus. Without crescentin, the cells adopt a straight-rod morphology. Crescentin has characteristic features of IF proteins including the ability to assemble into filaments in vitro without energy or cofactor requirements. In vivo, crescentin forms a helical structure that colocalizes with the inner cell curvatures beneath the cytoplasmic membrane. We propose that IF-like filaments of crescentin assemble into a helical structure, which by applying its geometry to the cell, generates a vibrioid or helical cell shape depending on the length of the cell.     

A growing family: the expanding universe of the bacterial cytoskeleton

Cytoskeletal proteins are important mediators of cellular organization in both eukaryotes and bacteria. In the past, cytoskeletal studies have largely focused on three major cytoskeletal families, namely the eukaryotic actin, tubulin, and intermediate filament (IF) proteins and their bacterial homologs MreB, FtsZ, and crescentin. However, mounting evidence suggests that these proteins represent only the tip of the iceberg, as the cellular cytoskeletal network is far more complex. In bacteria, each of MreB, FtsZ, and crescentin represents only one member of large families of diverse homologs. There are also newly identified bacterial cytoskeletal proteins with no eukaryotic homologs, such as WACA proteins and bactofilins. Furthermore, there are universally conserved proteins, such as the metabolic enzyme CtpS, that assemble into filamentous structures that can be repurposed for structural cytoskeletal functions. Recent studies have also identified an increasing number of eukaryotic cytoskeletal proteins that are unrelated to actin, tubulin, and IFs, such that expanding our understanding of cytoskeletal proteins is advancing the understanding of the cell biology of all organisms. Here, we summarize the recent explosion in the identification of new members of the bacterial cytoskeleton and describe a hypothesis for the evolution of the cytoskeleton from self-assembling enzymes.     

The bacterial cytoskeleton: in vivo dynamics of the actin-like protein Mbl of Bacillus subtilis

Mbl is a bacterial actin homolog that controls cell morphogenesis in Bacillus subtilis. A functional GFP-Mbl fusion protein was used to examine the behavior of the helical cables formed by Mbl protein in live B. subtilis cells. The cables undergo dynamic changes during cell cycle progression. They are stable but not rigid while elongating in parallel with cell growth, and they require septum formation to divide/cleave. Fluorescence recovery after photobleaching (FRAP) analysis showed that the cables are continuously remodeled during cell elongation. Turnover occurs along the length of the helical Mbl filaments, with no obvious polarity and a recovery half-time of about 8 min. These findings have important implications for the nature of bacterial cell wall architecture and synthesis.