Structural and dynamic states of actin in the erythrocyte

Analysis of the nucleotide tightly associated with isolated erythrocyte cytoskeletons show it to be ADP, rather then ATP. This confirms that at least a major part of the erythrocyte actin is in the F-form. A re-evaluation of the stoichiometry of spectrin and actin in the erythrocyte (taking account of a gross difference between the color responses of the two proteins on staining of electrophoretic gels) leads to values of 1x10(5) and 5x10(5) for the number of molecules of spectrin tetramer and actin respectively per cell. It has been found possible to perform spectrophotometric DNAase I assays fro actin on lysed whole cells. The concentration of monomeric actin at 0 degrees C is approximately 16 μg/ml packed cells. After washing the lysed cells the monomer pool is not re-established, indicating that only a small proportion of the actin subunits are free to dissociate. The actin monomer concentration in the cytosol remains unchanged after equilibration of the cells with cytochalasin E. The ability of actin-containing complexes in the membrane to nucleate the polymerization of added G-actin was measured fluorimetrically; it was found that membranes incubated with cytochalasin E were completely inert with respect to nucleating activity under conditions that favor appreciable growth at the slowly-growing (“pointed”) ends of free actin filaments. This suggests that these ends of the actin “protofilaments” in the red cell are blocked or sterically obstructed. After treatment of the membranes with guanidine hydrochloride under conditions that dissociate F-actin, the measured concentration of actin monomer rises to approximately 180 μg/ml of packed cells, which is nearly 70 percent of the total actin content. On treatment with trypsin in the presence of DNAase, the spectrin and 4.1 are extensively degraded, but the actin remains undamaged. This treatment, followed by exposure to guanidine hydrochloride, causes a further rise in the concentration of actin responsive to the DNAase assay to 250 μg/ml of cells, compared with 270 μg/ml estimated by densitometry of stained gels. The oligomeric complex, consisting of actin, spectrin, and 4.1, that is extracted from the membrane at low ionic strength, generates no detectable actin monomer after the same treatment. From literature data on the number of cytochalasin binding sites per cell and our value for the total actin content, we obtain a number-average degree of polymerization for actin in the membrane of 12-17. The results lead to a model for the structure of the cytoskeletal network and suggest some consequences of metabolic depletion.     

Water Flow Through Vessel Perforation Plates--The Effects of Plate Angle and Thickness for Liriodendron tulipifera

The effects of scalariform perforation plate thickness and anglewithin vessel elements of Liriodendron tulipifera were studiedwith a computational fluid dynamical model. The pressure gradientand hence resistance to flow through the plate increased asthe perforation plate increased in thickness. Increasing theangle of the plate relative to the axis of the vessel (samenumber of pores) also increased the pressure gradient alongthe modelled cell. For the model matching the actual vesselelement, the plate contributed 8% to the flow resistance ofthe vessel element. This contribution increased only to 11%for doubled plate thickness, and to 14% for a plate at an angleof 60° to the vessel axis. The perforation plate alteredthe velocity profile across the vessel element, but to a differentextent depending on the angle of the plate. A plate at an acuteangle to the vessel axis has little effect on the paraboloidprofile as found upstream from the plate, while obtuse angleplates change the flow profile such that fluid through poresnear the wall is accelerated to a greater velocity than foundin the centre of the cell. Key words: Conductance, perforation plate, vessel, water flow     

Organ specificity and transcriptional control of metabolic routes revealed by expression QTL profiling of source-sink tissues in a segregating potato population

Background  

With the completion of genome sequences belonging to some of the major crop plants, new challenges arise to utilize this data for crop improvement and increased food security. The field of genetical genomics has the potential to identify genes displaying heritable differential expression associated to important phenotypic traits. Here we describe the identification of expression QTLs (eQTLs) in two different potato tissues of a segregating potato population and query the potato genome sequence to differentiate between cis- and trans-acting eQTLs in relation to gene subfunctionalization.     

The topography of tip growth in a plant cell

Tips of young Phycomyces sporangiophores were dusted with starch grains, and growth photographically recorded. Rates of longitudinal displacement from the cell tip of individual markers were determined, also corresponding rates of change of cell diameter. From these the magnitude and spatial distribution of "relative elemental growth rates" along both longitudinal and circumferential axes of the cell were obtained. Growth rates in these two directions are functions of distance from the cell apex, and have different spatial distributions. In particular, rates of growth in cell circumference are complexly patterned. Relative elemental growth rates in length and in girth are approximately equal and maximal at the cell's apex, with a value of 2.4 mm. mm.^–1 hr.^–1. The characteristic shape of the tip is maintained constant in the face of its changing substance and position. This shape reflects a steady state of the cell's constituent growth patterns. At every point the growing membrane simultaneously expands in the two dimensions of its surface. The degree of polarization or directional preference of growth is measured by the ratio of longitudinal to circumferential relative elemental growth rate at any point. The ratio is not constant, but changes with position along the tip. This fact does not support the idea that membrane growth is based upon a quantal "growth event." Possible causal factors in oriented membrane growth are discussed.