The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts

The formation of a dynamic tubulovesicular membrane network that resembles the endoplasmic reticulum (ER) has been observed in extracts of cultured chick embryo fibroblasts (CEF cells) using video-enhanced differential interference contrast microscopy. Initially, membranes in the CEF extracts appeared amorphous and aggregated, but with time, membrane tubules moved out along stationary microtubules. The membrane tubules formed new branches on intersecting microtubules and fused with other branches to form a network of interconnected polygons. The tubulovesicular network was solubilized by detergent and took on a beaded morphology in a hypotonic buffer. Formation of the tubulovesicular network required ATP and microtubules. The network did not contain elements of the plasma membrane, Golgi apparatus, or mitochondria but could be labeled with ER markers. We suggest that the tubulovesicular network contains components from the ER and is formed by membrane associated motors moving upon microtubules in a process we call microtubule-dependent tethering.     

13C NMR studies of D- and L-phenylalanine binding to cobalt(II) carboxypeptidase A

13C NMR T1 and T2 measurements have been performed on cobalt(II) substituted carboxypeptidase A in the presence of carboxylate-13C-enriched L- and D-phenylalanine. Upon binding to the cobalt enzyme, the longitudinal and transverse relaxation rates T1p-1 and T2p-1 of these inhibitors are enhanced significantly compared to the zinc enzyme, allowing both determination of an affinity constant for inhibitor binding, K, and calculation of the metal-13C carboxylate distances. The L-and D- Phe concentration dependence of T2p-1 yields affinity constants of 290 +/- 60M-1 and 670 +/- 90M-1. The distance measurements calculated for Co-13C from T1p-1 are 0.39 +/- 0.04 and 0.42 +/- 0.04 nm for L-Phe and D-Phe. Both values are too great for direct coordination of their carboxylate groups to the metal atom. Upon formation of their respective ternary enzyme.Phe.N3- complexes, the distances are essentially unaltered. In conjunction with electronic absorption studies on these complexes it can be concluded that N3-, but not the amino acid carboxylate, is bound to the metal.     

Findings of new and rare fish species in the coastal zone of the Crimea (the Black Sea)

Characteristics of seven Atlantic-Mediterranean fish species are provided. Of them, four (gobies—yellow-headed Gobius xanthocephalus and red-mouthed G. cruentatus, incognito blenny Parablennius incognitus, and greater pipefish Syngnathus acus) were either first found in the Black Sea coastal zone of the Crimea, or their findings were doubted, and three species (thin-lipped grey mullet Liza ramada, European barracuda Sphyraena sphyraena, and Black Sea salmon Salmo trutta labrax) are very rare. Over the last decade, the authors recorded 15 fish species; of them, seven Atlantic-Mediterranean species have dispersed along the Crimean coasts by now, one endemic of the far eastern seas (chameleon goby Tridentiger trigonocephalus) has naturalized in Sevastopol Bay, four invading species, freshwater by origin, permanently occur in the estuary zones of the peninsula; the remaining three species are known from single findings and apparently belong to the category of accidental fish. An increase in the species composition of ichthyofauna off the Black Sea coasts of the Crimea results from the on-going process of mediterranization—the invasion in the estuary of freshwater tolerant species at the expense of accidental species that penetrated here either independently or with ballast waters.     

The fine structure of a microplate-microtubule array,microfilaments and polyhedral body associated microtubules in several species of Anabaena

A microplate-microtubule array was observed in Anabaena sp. (B-378). This structure consists of an arched plate, about 8 nm thick, and various microtubules, 12 nm in diameter and 50 nm long, arranged in rows. The microtubules project at right angles from one side of the plate into the cytoplasm or towards the plasma membrane. Up to twelve microplate-microtubule arrays were observed in a single section of a cell.Microfilaments, about 2.8 nm in diameter and of undetermined length, were observed in four isolates of Anabaena. The microfilaments were always found in bundles, which varied in size, up to 0.63 m across and 0.91 long.Microtubules, 10 nm in diameter and about 150 nm in length, were observed associated with one facet of polyhedral bodies in 8 out of 20 isolates of Anabaena. The microtubules occurred in groups of up to 20 or more, and were always oriented with the long axis parallel to a facet of a polyhedral body. In cross section, the microtubules had an electron transparent lumen 5 nm wide and a wall 2.5 nm thick.These structures are compared to previously deseribed microtubules and microfilaments.     

Effect of dimethyl sulfoxide (DMSO) on zinc availability (L-value), growth and metabolic activities of rice plants

Summary Effects of soil application (0.01, 0.1 and 1%) and foliar sprays (0.001, 0.01 and 0.1%) of dimethyl sulfoxide (DMSO) and soil application of Zn (10 and 20 ppm) on growth, yield, photosynthetic pigments and some enzymatic activities of rice (Oryza sativa L. variety Jaya) were investigated in a Zn-deficient soil under pot culture trials. Control plants showed typical Zn deficiency symptoms, very low dry matter and chlorophyll contents and significantly lower activities of carbonic anhydrase and tryptophan synthetase. Application of 10 or 20 ppm Zn to the soil markedly improved plant growth, chlorophyll content, enzymatic activities, Zn availability (L-value), and grain yield. Application of 1% DMSO to the soil proved to be severly phytotoxic for plant growth and dry matter accumulation. Application of lower doses of DMSO to the soil (0.01 and 0.1%) or as foliar sprays (0.001 and 0.01%), however, slightly increased dry weights of all plant parts at 45 days after transplanting. Grain yield was significantly increased by all DMSO treatments, except 1% soil application which completely suppressed the grain formation.Enzymatic activities and chlorophyll and carotenoid accumulation showed concentration-dependent stimulation or inhibition by DMSO treatments. Carbonic anhydrase activity was significantly increased by most of the DMSO treatments whereas tryptophan synthetase activity was stimulated only by the lowest dose of soil (0.01%) and foliar (0.001%) applications. Chlorophyll content was stimulated by lower doses of foliar application of DMSO (0.001 and 0.01%) but the other DMSO treatments brought about a concentration-dependent decrease in chlorophyll accumulation. Carotenoid accumulation was inversely related to that of chlorophylls. Chlorophyll: carotenoid ratio was increased by all DMSO and Zn treatments except 1% soil application of DMSO. Zn availability (L-value) of the soil was increased by all DMSO treatments. Zn content of leaf blades was positively correlated with chlorophyll content (r=0.579), tryptophan synthetase (r=0.700) and carbonic anhydrase (r=0.537) activities but negatively correlated with that of carotenoid content (r=–0.896). The possible mechanisms of DMSO on plants are discussed in relation to its possible agricultural utility.Publication No. 811 under Journal Series of the Experiment Station, G.B. Pant of Agriculture & Technology, Pantnagar (Nainital), India.     

Ecophysiological studies of Sonoran Desert plants

Summary The gas exchange of two Sonoran Desert plants was measured near optimum soil water conditions occurring in the summer and winter. Photosynthesis and stomatal conductance rates of a drought-deciduous shrub, Ambrosia deltoidea, and an evergreen non-riparian tree, Olneya tesota, are mainly affected by plant water potential. During such periods the diurnal gas exchange patterns are characterized by maximum rates of photosynthesis and stomatal conductance occurring early in the day, which decrease progressively thereafter. The effect of plant water potential on gas exchange is both direct and indirect. Decreasing plant water potential indirectly affects ¹⁴CO₂ assimilation by closure of stomata, and the effect is similar in both species. However, the direct effects of decreasing plant water potential are dissimilar in the two species. Plants of the shrub species have a higher potential maximum photosynthesis, but are more sensitive to plant water stress than are plants of the tree species. Both species respond to favorable growth conditions in the summer and winter, and have the potential for rapid carbon input into the Sonoran Desert ecosystem.This research was supported by National Science Foundation Grant BMS 74-02671-A04, through the U.S./I.B.P. Desert Biome     

An undescribed species of Drechslera (Dematiaceae) from India

Drechslera boeremae sp. nov. is described from India, having been isolated from the leaf-spot lesions of Portulaca oleracea L., a commonly grown vegetable.Contribution No. 555 from the Department of Mycology and Plant Pathology.     

Studies on a nonpolysomal ribonucleoprotein coding for myosin heavy chains from chick embryonic muscles.

A messenger ribonucleoprotein (mRNP) particle containing the mRNA coding for the myosin heavy chain (MHC mRNA) has been isolated from the postpolysomal fraction of homogenates of 14-day-old chick embryonic muscles. The mRNP sediments in sucrose gradient as 120 S and has a characteristic buoyant density of 1.415 g/cm3, which corresponds to an RNA:protein ratio of 1:3.8. The RNA isolated from the 120 S particle behaved like authentic MHC mRNA purified from chick embryonic muscles with respect to electrophoretic mobility and ability to program the synthesis of myosin heavy chain in a rabbit reticulocyte lysate system as judged by multi-step co-purification of the in vitro products with chick embryonic leg muscle myosin added as carrier. The RNA obtained from the 120 S particle was as effective as purified MHC mRNA in stimulating the synthesis of the complete myosin heavy chains in rabbit reticulocyte lysate under conditions where non-muscle mRNAs had no such effect. Analysis of the protein moieties of the 120 S particle by sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows the presence of seven distinct polypeptides with apparent molecular weights of 44,000, 49,000, 53,000, 81,000, 83,000, and 98,000, whereas typical ribosomal proteins are absent. These results indicate that the 120 S particles are distinct cellular entities unrelated to ribosomes or initiation complexes. The presence of muscle-specific mRNAs as cytoplasmic mRNPs suggests that these particles may be involved in translational control during myogenesis in embryonic muscles.     

Phage typing of cholera vibrios with different sets of phages]

A comparative study was made of two sets of Mukerjee and Drozhevkina-Arutyunova's bacteriophages in typing 514 strains of the El Tor vibrios and 45 strains of clasic biotype. It was shown that the Mukerjee or Drozhevkina-Arutyunova's phages could be used for the typing of cholera vibrios. The phages of the latter set prove to detect more phage types (18 against 11); they determine both the phage type and the biotype at the same time. The typing of cholera vibrios of both biotypes is possible, and the percentage of nontyping strains left is comparatively low (5.2 against 23.5 after Mukerjee). A table of the phage correspondence was made; it permits to obtain comparable data in using any set of the typing phages.