Saccharomyces cerevisiae Kelch Proteins and Bud14 Protein Form a Stable 520-kDa Formin Regulatory Complex That Controls Actin Cable Assembly and Cell Morphogenesis

Formins perform essential roles in actin assembly and organization in vivo, but they also require tight regulation of their activities to produce properly functioning actin structures. Saccharomyces cerevisiae Bud14 is one member of an emerging class of formin regulators that target the FH2 domain to inhibit actin polymerization, but little is known about how these regulators are themselves controlled in vivo. Kelch proteins are critical for cell polarity and morphogenesis in a wide range of organisms, but their mechanistic roles in these processes are still largely undefined. Here, we report that S. cerevisiae Kelch proteins, Kel1 and Kel2, associate with Bud14 in cell extracts to form a stable 520-kDa complex with an apparent stoichiometry of 2:2:1 Bud14/Kel1/Kel2. Using pairwise combinations of GFP- and red fluorescent protein-tagged proteins, we show that Kel1, Kel2, and Bud14 interdependently co-localize at polarity sites. By analyzing single, double, and triple mutants, we show that Kel1 and Kel2 function in the same pathway as Bud14 in regulating Bnr1-mediated actin cable formation. Loss of any component of the complex results in long, bent, and hyper-stable actin cables, accompanied by defects in secretory vesicle traffic during polarized growth and septum formation during cytokinesis. These observations directly link S. cerevisiae Kelch proteins to the control of formin activity, and together with previous observations made for S. pombe homologues tea1p and tea3p, they have broad implications for understanding Kelch function in other systems.     

Integrating cellular dimensions with cell differentiation during early development

Early embryo development is characterized by alteration of cellular dimensions and fating of blastomeres. An emerging concept is that cell size and shape drive cellular differentiation during early embryogenesis in a variety of model organisms. In this review, we summarize recent advances that elucidate the contribution of the physical dimensions of a cell to major embryonic transitions and cell fate specification in vivo. We also highlight techniques and newly evolving methods for manipulating the sizes and shapes of cells and whole embryos in situ and ex vivo. Finally, we provide an outlook for addressing fundamental questions in the field and more broadly uncovering how changes to cell size control decision making in a variety of biological contexts.     

Dual function of Yap in the regulation of lens progenitor cells and cellular polarity

Hippo-Yap signaling has been implicated in organ size determination via its regulation of cell proliferation, growth and apoptosis (Pan, 2007). The vertebrate lens comprises only two major cell types, lens progenitors and differentiated fiber cells, thereby providing a relatively simple system for studying size-controlling mechanisms. In order to investigate the role of Hippo-Yap signaling in lens size regulation, we conditionally ablated Yap in the developing mouse lens. Lens progenitor-specific deletion of Yap led to near obliteration of the lens primarily due to hypocellularity in the lens epithelium (LE) and accompanying lens fiber (LF) defects. A significantly reduced LE progenitor pool resulted mainly from failed self-renewal and increased apoptosis. Additionally, Yap-deficient lens progenitor cells precociously exited the cell cycle and expressed the LF marker, β-Crystallin. The mutant progenitor cells also exhibited multiple cellular and subcellular alterations including cell and nuclear shape change, organellar polarity disruption, and disorganized apical polarity complex and junction proteins such as Crumbs, Pals1, Par3 and ZO-1. Yap-deficient LF cells failed to anchor to the overlying LE layer, impairing their normal elongation and packaging. Furthermore, our localization study results suggest that, in the developing LE, Yap participates in the cell context-dependent transition from the proliferative to differentiation-competent state by integrating cell density information. Taken together, our results shed new light on Yap's indispensable and novel organizing role in mammalian organ size control by coordinating multiple events including cell proliferation, differentiation, and polarity.     

水文尺度转换探讨

水文尺度问题是当今水文学研究的热点和前沿.水文尺度包括过程尺度、观测尺度和模拟尺度三方面含义.主导过程尺度作为尺度的特征量,是水文研究中的重点.水文尺度转换工作包括了对水文模式、参数、状态变量和输入的尺度转换四方面,每一方面都有其特殊的研究方法.水文系统的有组织复杂性、时空变异性和数据不足是当前水文研究的难点所在.自然河网的自相似特性使其成为水文尺度研究的重要组成部分.进一步深入研究水文尺度问题需要多种研究思路、技术手段和理论工具的结合.     

Cryptic individual scaling relationships and the evolution of morphological scaling

Morphological scaling relationships between organ and body size—also known as allometries—describe the shape of a species, and the evolution of such scaling relationships is central to the generation of morphological diversity. Despite extensive modeling and empirical tests, however, the modes of selection that generate changes in scaling remain largely unknown. Here, we mathematically model the evolution of the group‐level scaling as an emergent property of individual‐level variation in the developmental mechanisms that regulate trait and body size. We show that these mechanisms generate a “cryptic individual scaling relationship” unique to each genotype in a population, which determines body and trait size expressed by each individual, depending on developmental nutrition. We find that populations may have identical population‐level allometries but very different underlying patterns of cryptic individual scaling relationships. Consequently, two populations with apparently the same morphological scaling relationship may respond very differently to the same form of selection. By focusing on the developmental mechanisms that regulate trait size and the patterns of cryptic individual scaling relationships they produce, our approach reveals the forms of selection that should be most effective in altering morphological scaling, and directs researcher attention on the actual, hitherto overlooked, targets of selection.     

Statistical shape modelling versus linear scaling: Effects on predictions of hip joint centre location and muscle moment arms in people with hip osteoarthritis

Marker-based dynamic functional or regression methods are used to compute joint centre locations that can be used to improve linear scaling of the pelvis in musculoskeletal models, although large errors have been reported using these methods. This study aimed to investigate if statistical shape models could improve prediction of the hip joint centre (HJC) location. The inclusion of complete pelvis imaging data from computed tomography (CT) was also explored to determine if free-form deformation techniques could further improve HJC estimates. Mean Euclidean distance errors were calculated between HJC from CT and estimates from shape modelling methods, and functional- and regression-based linear scaling approaches. The HJC of a generic musculoskeletal model was also perturbed to compute the root-mean squared error (RMSE) of the hip muscle moment arms between the reference HJC obtained from CT and the different scaling methods. Shape modelling without medical imaging data significantly reduced HJC location error estimates (11.4 ± 3.3 mm) compared to functional (36.9 ± 17.5 mm, p = <0.001) and regression (31.2 ± 15 mm, p = <0.001) methods. The addition of complete pelvis imaging data to the shape modelling workflow further reduced HJC error estimates compared to no imaging (6.6 ± 3.1 mm, p = 0.002). Average RMSE were greatest for the hip flexor and extensor muscle groups using the functional (16.71 mm and 8.87 mm respectively) and regression methods (16.15 mm and 9.97 mm respectively). The effects on moment-arms were less substantial for the shape modelling methods, ranging from 0.05 to 3.2 mm. Shape modelling methods improved HJC location and muscle moment-arm estimates compared to linear scaling of musculoskeletal models in patients with hip osteoarthritis.     

Closing the loops: new insights into the role and regulation of actin during cell polarization

In this review, we summarize recent results on the understanding of actin organization and cell polarization with an emphasis on the critical role of actin during this process. We first report on the advances made in understanding the function and mechanism of formin family proteins in the nucleation of actin filaments. We also discuss how formins and other regulators of actin dynamics are thought to be involved in the generation of cell polarity. In the second part we discuss new findings indicating that, rather than using a linear pathway from signal transduction to cytoskeleton re-organization, cell polarity is established through bidirectional interplay between these processes. We describe the various types of feedback loops identified and point out common schemes. Finally we briefly summarize the emerging role of actinlike proteins in the generation of polarity in prokaryotes that implies an early origin of actin's role in cell polarity.     

Uncovering principles that control septin-septin interactions

Septins comprise a conserved family of GTPases important in cytokinesis. These proteins polymerize into filaments from rod-shaped heteromeric septin complexes. Septins interact with one another at two interfaces (NC and G) that alternate within the complex. Here, we show that small mutations at the N terminus greatly enhance the formation of SEPT2 homopolymers. Taking advantage of this mutation to examine polymer formation using SEPT2 alone, we show that both NC and G interfaces are required for filament formation. However, co-expression of wild type SEPT2 with SEPT2 containing mutations at either NC or G interfaces revealed that only the NC mutant suppressed filament formation. NC mutants are able to interact with one another at putative G interfaces, whereas G mutants fail to interact at NC interfaces. In addition, all promiscuous septin pairwise interactions occur at the G interface. These findings suggest that G interface interactions must occur before NC interactions during polymer formation.     

The formation and patterning of leaves: recent advances

Leaves, the plants major photosynthetic organs, form through the activity of groups of pluripotent cells, termed shoot apical meristems (SAMs), located at the growing tips of plants. Leaves develop with a dorso–ventral asymmetry, with the adaxial surface adjacent to the meristem and the abaxial surface developing at a distance from it. Molecular genetic studies have shown that the correct specification of adaxial/abaxial polarity requires communication between the incipient leaf and the meristem, and that the juxtaposition of adaxial/abaxial fates is necessary for lamina outgrowth (Waites and Hudson 1995; McConnell et al. 2001). Over the last few years, a number of factors that control cell fate specification in the apex have been identified. This review will focus on recent advances on distinct but overlapping aspects of leaf development, namely, the transition from meristem to leaf fate and the specification of abaxial/adaxial polarity and its possible role in leaf growth.     

The effectiveness of scaling procedures for comparing ground reaction forces

Various scaling methods are used when attempting to remove the influence of anthropometric differences on ground reaction forces (GRF) when comparing groups. Though commonly used, ratio scaling often results in an over-correction. Allometric scaling has previously been suggested for kinetic variables but its effectiveness in partialing out the effect of anthropometrics is unknown due to a lack of consistent application. This study examined the effectiveness of allometric scaling vertical, braking and propulsive GRF and loading rate for 84 males and 47 females while running at 4.0 m/s. Raw, unfiltered data were ratio scaled by body mass (BM), height (HT), and BM multiplied by HT (BM1HT). Gender specific exponents for allometric scaling were determined by performing a log-linear (for BM and HT individually) or log-multilinear regression (BMHT). Pearson productmoment correlations were used to assess the effectiveness of each scaling method. Ratio scaling by BM, HT, or BM1HT resulted in an over-correction of the data for most variables and left a considerable portion of the variance still attributable to anthropometrics. Allometric scaling by BM successfully removed the effect of BM and HT for all variables except for braking GRF in males and vertical GRF in females. However, allometric scaling for BMHT successfully removed the effect of BM and HT for all reactionary forces in both genders. Based on these results, allometric scaling for BMHT was the most appropriate scaling method for partialing out the effect of BM and HT on kinetic variables to allow for effective comparisons between groups or individuals.     

细胞壁在细胞极性建立和胚胎发生中的作用

植物细胞壁是一个活性的动态结构,其结构层次与组分随着发育进程而发生变化,且广泛参与细胞的各项生命活动,特别是在参与细胞命运决定、充当细胞发育信使、调控植物胚胎早期极性建立以及模式建成等方面发挥重要作用。     

A biomechanical analysis of mandibular scaling in old world monkeys

Scaling of mandibular dimensions in male Old World monkeys was investigated. Mandibular condyle length, width, and area were regressed separately against body mass and mandibular length for a total of 14 species of Cercopithecoids. Scaling of mandibular depth and width against both body mass and mandibular length were also investigated. When results of regression analysis using the two different independent variables (body mass and mandibular length) were compared, there were significant pattern differences in scaling of cercopithecines versus colobines. Compared to body mass, male cercopithecines had relatively large mandibles (length, width, and depth) and also relatively large condyles (length, width, and area). However, compared to mandibular length, cercopithecines had relatively transversely thin and shallow mandibles and relatively narrow condyles. It is shown that a “biomechanical” interpretation of mandibular scaling patterns against body mass in Old World monkeys demonstrates only that cercopithecines have prognathic faces, an already well-known and well-documented condition. When the biomechanical effects of prognathic faces are controlled for (by scaling against mandibular length), it is shown that cercopithecines possess special adaptations in condyle length while colobines possess special adaptations in condyle width and mandibular depth and width. These results clearly demonstrate the importance of selecting a relevant reference variable in scaling studies where biomechanical interpretations are attempted.     

Shape and size changes induced by taurine depletion in neonatal cardiomyocytes

Summary Taurine is a very important organic osmolyte in most adult cells. Because of this property it has been proposed that large changes in the intracellular content of taurine can osmotically stress the cell, causing changes in its size and shape. This hypothesis was examined by measuring cell dimensions of taurine deficient cardiomyocytes using confocal microscopy. Incubation of isolated neonatal rat myocytes with medium containing 5mM-alanine led to a 55% decrease in intracellular taurine content. Associated with the loss of taurine was a reduction in cell size. Two factors contributed to the change in cell size. First, there was a shift in cell shape, favoring the smaller of the two cellular configurations commonly found in the myocyte cell culture. Second, the size of the polyhedral configuration was reduced after ßalanine treatment. These same two events also contributed to size reduction in cardiomyocytes incubated with medium containing 30mM mannitol. Nonetheless, some qualitative differences exist between cells osmotically stressed by increasing the osmolality of the incubation medium and decreasing intracellular osmolality. The results support a role for taurine in the regulation of osmotic balance in the neonatal cardiomyocyte.     

Whole-cell modeling framework in which biochemical dynamics impact aspects of cellular geometry

A mathematical framework for modeling biological cells from a physicochemical perspective is described. Cells modeled within this framework consist of at least two regions, including a cytosolic volume encapsulated by a membrane surface. The cytosol is viewed as a well-stirred chemical reactor capable of changing volume while the membrane is assumed to be an oriented 2-D surface capable of changing surface area. Two physical properties of the cell, namely volume and surface area, are determined by (and determine) the reaction dynamics generated from a set of chemical reactions designed to be occurring in the cell. This framework allows the modeling of complex cellular behaviors, including self-replication. This capability is illustrated by constructing two self-replicating prototypical whole-cell models. One protocell was designed to be of minimal complexity; the other to incorporate a previously reported well-known mechanism of the eukaryotic cell cycle. In both cases, self-replicative behavior was achieved by seeking stable physically possible oscillations in concentrations and surface-to-volume ratio, and by synchronizing the period of such oscillations to the doubling of cytosolic volume and membrane surface area. Rather than being enforced externally or artificially, growth and division occur naturally as a consequence of the assumed chemical mechanism operating within the framework.     

The human small glutamine-rich TPR-containing protein is required for progress through cell division

Eukaryotic organisms from yeast to human harbor genes encoding the small glutamine-rich tetratricopeptide repeat-containing (SGT) protein. Work presented here demonstrated the presence of human SGT (hSGT) protein in a panel of human cell lines and throughout the cell cycle. To identify cellular processes in which hSGT is involved, knock down populations were analyzed which were generated through transfection of hsgt-specific small interfering RNA. Most strikingly, depletion of hSGT led to reduced proliferation of the affected cell populations while the mitotic index was increased. Time-lapse video microscopy revealed that cells from hSGT-depleted populations were unable to complete cell division due to mitotic arrest which was frequently followed by cell death. Further evidence for a role in cell division was given by the accumulation of hSGT in the midzone and the midbody, and by a mitosis-specific migration pattern of hSGT as detected by Western blotting after SDS-PAGE or two-dimensional gel electrophoresis. In conclusion, results obtained in this study demonstrate that hSGT protein is a constitutive component of all human cell lines tested and that this protein is essential for successful completion of cell division.     

The contractile ring

Summary Techniques of individual cell selection and precise ultramicrotomy have been employed to demonstrate that the contractile ring of cleaving HeLa cells is a transitory cytoplasmic organelle of distinctive fine structure and location. The contractile ring is an uninterrupted annulus encircling the equator of dividing cells exactly where the cleavage furrow forms. It is about 10 microns wide, up to 0.2 microns in thickness, and is composed exclusively of circumferentially aligned thin filaments 40–70 Å in diameter. Contractile ring filaments appear to be associated with the overlying plasma membrane.Controlled experiments with a mold metabolite (cytochalasin B) reveals that within a few minutes the drug abolishes the ability of HeLa cells to undergo cytokinesis. Cytochalasin B seems to decompose the contractile ring. It has no other clearly identifiable effects on other cell structures, notably the mitotic apparatus. Cytochalasin B is the only drug known which selectively inhibits cytokinesis in animal cells.In conclusion, the contractile ring is the most likely organelle responsible for cytokinesis in HeLa cells. Similar organelles probably occur in all cleaving animal cells. Successful cleavage depends upon the structural and functional integrity of the contractile ring.