How do cytokinins affect the cell?

The lecture presents modern knowledge of the mechanisms of cytokinin perception and signal transduction to the genes of primary and secondary responses. It also demonstrates the relations between the rapid cytokinin-induced processes and cytokinin-induced physiological effects. The characteristics of the cytokinin regulatory system and its role in the control of plant growth and development are discussed.     

How do prokaryotic cells cycle?

This issue of Current Biology features five reviews covering various key aspects of the eukaryotic cell cycle. The topics include initiation of chromosome replication, assembly of the mitotic spindle, cytokinesis, the regulation of cell-cycle progression, and cell-cycle modeling, focusing mainly on budding yeast, fission yeast and animal cell model systems. The reviews underscore common themes as well as key differences in the way these processes are carried out and regulated among the different model organisms. Consequently, an important question is how cell-cycle mechanisms and controls have evolved, particularly in the broader perspective of the three domains of life.     

How do resident stem cells repair the damaged myocardium?

It has been a decade since the monumental discovery of resident stem cells in the mammalian heart, and the following studies witnessed the continuous turnover of cardiomyocytes and vascular cells, maintaining the homeostasis of the organ. Recently, the autologous administration of c-kit-positive cardiac stem cells in patients with ischemic heart failure has led to an incredible outcome; the left ventricular ejection fraction of the celltreated group improved from 30% at the baseline to 38% after one year and to 42% after two years of cell injection. The potential underlying mechanisms, before and after cell infusion, are explored and discussed in this article. Some of them are related to the intrinsic property of the resident stem cells, such as direct differentiation, paracrine action, and immunomodulatory function, whereas others involve environmental factors, leading to cellular reverse remodeling and to the natural selection of "juvenile" cells. It has now been demonstrated that cardiac stem cells for therapeutic purposes can be prepared from tiny biopsied specimens of the failing heart as well as from frozen tissues, which may remarkably expand the repertoire of the strategy against various cardiovascular disorders, including non-ischemic cardiomyopathy and congenital heart diseases. Further translational investigations are needed to explore these possibilities.     

How do plant cell walls extend?

This article briefly summarizes recent work that identifies the biophysical and biochemical processes that give rise to the extension of plant cell walls. I begin with the biophysical notion of stress relaxation of the wall and follow with recent studies of wall enzymes thought to catalyze wall extension and relaxation. Readers should refer to detailed reviews for more comprehensive discussion of earlier literature (Taiz, 1984; Carpita and Gibeaut, 1993; Cosgrove, 1993).     

How do cells move along surfaces?

The movement of cells along surfaces is a complex phenomenon that consists of several interrelated processes, including cell-substratum adhesion, and extension and retraction of the cell edge, in which the actin cytoskeleton plays a crucial role. The past decade has seen increasingly detailed molecular-based investigations into cell motility, but it is still not known how molecular events are integrated to give cell movement. Molecular studies are now beginning to be linked to a more global concept of how whole cells move, and this combined approach promises to yield new insights into cell locomotion.     

How do microtubules guide migrating cells?

Microtubules have long been implicated in the polarization of migrating cells, but how they carry out this role is unclear. Here, we propose that microtubules determine cell polarity by modulating the pattern of adhesions that a cell develops with the underlying matrix, through focal inhibitions of contractility.     

How do yeast cells sense glucose?

A glucose-sensing mechanism has been described in Saccharomyces cerevisiae that regulates expression of glucose transporter genes. The sensor proteins Snf3 and Rgt2 are homologous to the transporters they regulate. Snf3 and Rgt2 are integral plasma membrane proteins with unique carboxy-terminal domains that are predicted to be localized in the cytoplasm. In a recent paper Ozcan and colleagues [Ozcan S, et al. EMBO J 1998; 17:2556-2773 (Ref. 1)] present evidence that the cytoplasmic domains of Snf3 and Rgt2 are required to transmit a glucose signal. They provide additional evidence to support their earlier assertion [Ozcan S, et al. Proc Natl Acad Sci USA 1996;93:12428-12432 (Ref. 2)] that glucose transport via Snf3 and Rgt2 is not involved in glucose sensing but, rather, that these proteins behave like glucose receptors. Other examples of transporter homologs with regulatory functions have recently been described in fungi as well [Madi L, et al. Genetics 1997; 146:499-508 (Ref. 3). and Didion T, et al. Mol Microbiol 1998;27:643-650 (Ref. 4)]. The identification of this class of nutrient sensors is an important step in elucidating the complex of regulatory mechanisms that leads to adaptation of fungi to different environments.     

How do so few control so many?

The separation of sister chromatids at the metaphase-to-anaphase transition is triggered by a protease called separase that is activated by the destruction of an inhibitory chaperone (securin). This process is mediated by a ubiquitin protein ligase called the anaphase-promoting complex or cyclosome (APC/C), along with a protein called Cdc20. It is vital that separase not be activated before every single chromosome has been aligned on the mitotic spindle. Kinetochores that have not yet attached to microtubules catalyze the sequestration of Cdc20 by an inhibitor called Mad2. Recent experiments shed important insight into how Mad2 molecules bound to centromeres through their association with a protein called Mad1 might be transferred to Cdc20 and thereby inhibit securin's destruction.     

How do cell walls regulate plant growth?

The cell wall of growing plant tissues has frequently been interpreted in terms of inextensible cellulose microfibrils 'tethered' by hemicellulose polymers attached to the microfibril surface by hydrogen bonds, with growth occurring when tethers are broken or 'peeled' off the microfibril surface by expansins. This has sometimes been described as the 'sticky network' model. In this paper, a number of theoretical difficulties with this model, and discrepancies between predicted behaviour and observations by a number of researchers, are noted. (i) Predictions of cell wall moduli, based upon the sticky network model, suggest that the cell wall should be much weaker than is observed. (ii) The maximum hydrogen bond energy between tethers and microfibrils is less than the work done in expansion and therefore breakage of such hydrogen bonds is unlikely to limit growth. (iii) Composites of bacterial cellulose with xyloglucan are weaker than pellicles of pure cellulose so that it seems unlikely that hemicelluloses bind the microfibrils together. (iv) Calcium chelators promote creep of plant material in a similar way to expansins. (v) Reduced relative 'permittivities' inhibit the contraction of cell wall material when an applied stress is decreased. Revisions of the sticky network model that might address these issues are considered, as are alternatives including a model of cell wall biophysics in which cell wall polymers act as 'scaffolds' to regulate the space available for microfibril movement. Experiments that support the latter hypothesis, by demonstrating that reducing cell wall free volume decreases extensibility, are briefly described.     

A genome-wide screen identifies conserved protein hubs required for cadherin-mediated cell–cell adhesion

Cadherins and associated catenins provide an important structural interface between neighboring cells, the actin cytoskeleton, and intracellular signaling pathways in a variety of cell types throughout the Metazoa. However, the full inventory of the proteins and pathways required for cadherin-mediated adhesion has not been established. To this end, we completed a genome-wide (∼14,000 genes) ribonucleic acid interference (RNAi) screen that targeted Ca²⁺-dependent adhesion in DE-cadherin–expressing Drosophila melanogaster S2 cells in suspension culture. This novel screen eliminated Ca²⁺-independent cell–cell adhesion, integrin-based adhesion, cell spreading, and cell migration. We identified 17 interconnected regulatory hubs, based on protein functions and protein–protein interactions that regulate the levels of the core cadherin–catenin complex and coordinate cadherin-mediated cell–cell adhesion. Representative proteins from these hubs were analyzed further in Drosophila oogenesis, using targeted germline RNAi, and adhesion was analyzed in Madin–Darby canine kidney mammalian epithelial cell–cell adhesion. These experiments reveal roles for a diversity of cellular pathways that are required for cadherin function in Metazoa, including cytoskeleton organization, cell–substrate interactions, and nuclear and cytoplasmic signaling.     

Boron deficiency: How does the defect in cell wall damage the cells?

Boron (B) is an essential micronutrient for vascular plants. Boron plays a structural role in cell walls through binding to pectic polysaccharides. It still remains unclear how B deficiency, and hence probably alterations in cell wall structure, leads to various metabolic disorders and cell death. To understand the process, we analyzed the physiological changes in suspension- cultured tobacco (Nicotiana tabacum) BY-2 cells under B deficiency. The results indicated that the cells deprived of B did not undergo a typical programmed cell death process. Oxidative damage was proven to be the direct and major cause of cell death. We discuss possible mechanisms for the generation and accumulation of reactive oxygen species under B deprivation.Key words: boron deficiency, cell death, cell wall, oxidative damage, pectic polysaccharides, rhamnogalacturonan II, tobaccoBoron (B) deficiency is the most widespread micronutrient deficiency around the world and causes large losses in crop production both quantitatively and qualitatively.¹ Boron deficiency affects vegetative and reproductive growth of plants resulting in inhibition of cell expansion, death of meristem and reduced fertility.²Plants contain B both in a water-soluble and insoluble form. In intact plants, the amount of water-soluble B fluctuates with the quantity of B supplied, while insoluble B does not.³ The appearance of B deficiency symptoms coincides with the decrease of water-insoluble B, from which it is concluded that the insoluble B is the functional form while the soluble B represents the surplus. We found at least 98% of the insoluble B in tobacco cells bound to the cell wall,⁴ and identified their molecular entity as the borate diester with rhamnogalacturonan II (RG-II) regions of pectic polysaccharides.⁵ The diester crosslinks pectic polysaccharides to form a network and thereby contributes to construction of a supramolecular cell wall structure.⁶ Mutant plants with altered RG-II structures are dwarf and sterile, indicating that the B-RG-II complex is essential for normal plant growth and development.⁷ Increasing evidence indicates that B is also essential for animals.⁸ The requirement for B in organisms lacking cell walls implies that B may also have additional roles in plants. To date, however, no molecule other than apiosyl residues in pectic polysaccharides has been demonstrated to form a borate ester which could be stable enough under physiological conditions. Thus it is reasonable to consider that B functions primarily, if not exclusively, as a structural component of the cell wall, and B deficiency symptoms arise from disturbance of the cell wall structure. How, then, does the disturbed cell wall structure lead to the damage and cell death that are observed under B deficiency? To understand the linkage, we have analyzed physiological changes of suspension-cultured tobacco (Nicotiana tabacum) BY-2 cells under B deficiency.When cells at the log phase of growth were transferred to B-free media, cell death was detectable as early as 12 h after the treatment. As cell walls play pivotal roles in plant development and growth, we assumed that the B deprivation, which probably causes aberrant cell wall structure, might induce programmed cell death (PCD) as an active response to eliminate damaged cells. Then we examined if the known biochemical hallmark of PCD could be observed in cells deprived of B (hereafter referred to as -B cells). However, internucleosomal DNA fragmentation, decrease in antioxidant content and antioxidant enzyme expression,⁹ or protection from death by cycloheximide, were not detected in these cells, suggesting that the cell death is necrosis. We found oxidative damage to be the direct and major cause of cell death, because -B cells contained more reactive oxygen species (ROS) than control cells, and because cell death was effectively suppressed by supplementing the media with lipophilic antioxidants. The deprivation treatment did not induce an oxidative burst, as the extracellular H₂O₂ concentration was not significantly different between -B and control cells at all time points examined. Resupply of B immediately suppressed cell death. Collectively, these results suggest that low but persistent ROS production occurred under the -B condition.In the study described above, we demonstrated that B deprivation, and hence probably a defective cell wall structure, leads to oxidative damage. How and why B deprivation induces ROS overproduction remains to be clarified. We hypothesize that ROS are originally produced as a signal for disturbance of the cell wall structure, and build up to a toxic level unless B is resupplied and the cell wall structure is restored. It has been reported that the mechanical strength of the squash root cell wall decreases within minutes after B deprivation.^10,11 The mechanical change could be brought about by insufficient crosslinking of pectic polysaccharides at RG-II regions, as the B-RG-II complex significantly contributes to the wall tensile strength.¹² If the cell wall becomes weaker and less resistant to turgor, then the plasma membrane would stretch. The change may lead to opening of mechanosensitive channels¹³ and generation of signals for the altered cell wall structure. To test this hypothesis, we are now analyzing the immediate and early responses of tobacco BY-2 cells to B deprivation, and preliminary results do indicate the involvement of Ca²⁺ influx in the responses. Identification of the mechanism by which cells sense the external B status will greatly contribute to our understanding of the cell wall-symplast interaction in plants.¹⁴     

How do African game animals control trypanosome infections?

African trypanosomiasis is endemic over much of sub-saharan Africa. But whereas domestic animals - especially cattle - often succumb to the infection, wild mammals generally show a high degree o f resistance. Many species o f wildlife living in tsetse-infested areas carry trypanosome infections, and so act as important reservoir hosts, but generally show no obvious ill-effects. How they survive the infection is an important question in understanding mechanisms o f trypanotolerance, yet relevant data are sparse and scattered. Here, Ayub Mulla and Roy Rickman review some of the previous studies to illustrate how complex the question is.     

How do tenascins influence the birth and life of a malignant cell?

Tenascins are large glycoproteins found in embryonic and adult extracellular matrices. Of the four family members, two have been shown to be overexpressed in the microenvironment of solid tumours: tenascin-C and tenascin-W. The regular presence of these proteins in tumours suggests a role in tumourigenesis, which has been investigated intensively for tenascin-C and recently for tenascin-W as well. In this review, we follow a malignant cell starting from its birth through its potential metastatic journey and describe how tenascin-C and tenascin-W contribute to these successive steps of tumourigenesis. We consider the importance of the mechanical aspect in tenascin signalling. Furthermore, we discuss studies describing tenascin-C as an important component of stem cell niches and present examples reporting its role in cancer therapy resistance.     

How do plants feel the heat?

In plants, the heat stress response (HSR) is highly conserved and involves multiple pathways, regulatory networks and cellular compartments. At least four putative sensors have recently been proposed to trigger the HSR. They include a plasma membrane channel that initiates an inward calcium flux, a histone sensor in the nucleus, and two unfolded protein sensors in the endoplasmic reticulum and the cytosol. Each of these putative sensors is thought to activate a similar set of HSR genes leading to enhanced thermotolerance, but the relationship between the different pathways and their hierarchical order is unclear. In this review, we explore the possible involvement of different thermosensors in the plant response to warming and heat stress.     

Down-regulation of Pkd2 by siRNAs suppresses cell–cell adhesion in the mouse melanoma cells

The Pkd2 gene encodes an integral protein (~130 kDa), named polycystin-2 (PC-2). PC-2 is mainly involved in autosomal dominant polycystic kidney disease. Recently, polycystin-1/polycystin-2 complex has been shown to act as an adhesion complex mediating or regulating cell–cell or cell–matrix adhesion, suggesting that PC-2 may play a role in cell–cell/cell–matrix interactions. Here, we knocked down the expression of Pkd2 gene with small interfering RNAs (siRNAs) in the mouse melanoma cells (B16 cells), indicating that the cells transfected with the targeted siRNAs significantly suppressed cell–cell adhesion, but not cell–matrix adhesion, compared to the cells transfected with non-targeted control (NC) siRNA. This study provides the first directly functional evidence that PC-2 mediates cell–cell adhesion. Furthermore, we demonstrated that PC-2 modulated cell–cell adhesion may be, at least partially, associated with E-cadherin. Collectively, these findings for the first time showed that PC-2 may mediate cell–cell adhesion, at least partially, through E-cadherin.