Phosphatidic Acid-Dependent Phosphorylation of a 29-kDa Protein by Protein Kinase Cα in Bovine Brain Cytosol

Abstract: Activation of phospholipase D (PLD) is involved in receptor-mediated signal transduction responses. Signaling from PLD to a downstream molecule(s) appears to be mediated by the PLD product phosphatidic acid (PA). A target molecule(s) of PA, however, has not yet been identified. The present study sought to define such a target molecule(s) of PA. In bovine brain cytosol, proteins with apparent molecular weights of 29,000 (p29) and 32,000 (p32) were prominently phosphorylated in the presence of PA, but not in its absence, indicating that there is a PA-regulated protein kinase (PARK) in bovine brain that phosphorylates p29 and p32. One of these substrates, p29, was purified to near homogeneity. Its partial amino acid sequence was determined and found to be identical to that of a known brain-specific 25-kDa protein (p25). The purified p29 was also readily recognized by and immunoprecipitated with an anti-p25 antibody. These results suggest that p29 is very similar to or identical with p25. Using the purified p29 as a substrate, PARK was purified to near homogeneity. The purified PARK had an apparent molecular weight of 80,000, was strongly recognized by an anti-protein kinase C (PKC)α antibody, and was activated by phosphatidylserine (PS) as well as PA. The PA- and PS-stimulated PARK activity was extremely augmented by the presence of 1 µM free Ca²⁺. In the presence of 1 mM EGTA, phorbol 12-myristate 13-acetate activated PARK synergistically with PA or PS. Similar results were obtained with the purified recombinant PKCα. From these results, it is suggested that the PARK activity purified might be attributed to PKCα. In p25-depleted bovine brain cytosol, which was prepared by treatment of bovine brain cytosol with the anti-p25 antibody, PA-dependent phosphorylation of p29, but not p32, was almost completely eliminated. When PKCα in bovine brain cytosol was depleted by its precipitation with the anti-PKCα antibody, neither p29 nor p32 in this PKCα-depleted cytosol was phosphorylated in the presence of PA. These results indicate that in bovine brain cytosol PA activates PKCα, which, in turn, phosphorylates p29, which may be identical with p25.     

Elevated CO2, rhizosphere processes,and soil organic matter decomposition

The rhizosphere is one of the key fine-scale components of C cycles. This study was undertaken to improve understanding of the potential effects of atmospheric CO2 increase on rhizosphere processes. Using C isotope techniques, we found that elevated atmospheric CO2 significantly increased wheat plant growth, dry mass accumulation, rhizosphere respiration, and soluble C concentrations in the rhizosphere. When plants were grown under elevated CO2 concentration, soluble C concentration in the rhizosphere increased by approximately 60%. The degree of elevated CO2 enhancement on rhizosphere respiration was much higher than on root biomass. Averaged between the two nitrogen treatments and compared with the ambient CO2 treatment, wheat rhizosphere respiration rate increased 60% and root biomass only increased 26% under the elevated CO2 treatment. These results indicated that elevated atmospheric CO2 in a wheat-soil system significantly increased substrate input to the rhizosphere due to both increased root growth and increased root activities per unit of roots. Nitrogen treatments changed the effect of elevated CO2 on soil organic matter decomposition. Elevated CO2 increased soil organic matter decomposition (22%) in the nitrogen-added treatment but decreased soil organic matter decomposition (18%) without nitrogen addition. Soil nitrogen status was therefore found to be important in determining the directions of the effect of elevated CO2 on soil organic matter decomposition.     

Accumulation of β-tubulin-containing fibres in the cortical domain ofSaccharomyces cerevisiae spheroplasts

Summary The distribution of microtubules inSaccharomyces cerevisiae was studied with the monoclonal antibody (mab) TU-14 against -tubulin. Immunoblotting and immunoprecipitation experiments with a strain overexpressing Tub2p confirmed that the mab TU-14 specifically recognized Tub2p. By immunofluorescence microscopy, the mab TU-14 attached to all known tubulin structures labelled with the standard polyclonal anti--tubulin antibody 206-1. In addition, the mab TU-14 revealed cortical patches in wild-type cells and an abundant network of fibres in the cortex of spheroplasts cultivated in nutrient medium. These cortical fibres seemed to be specific to spheroplasts and suggest that the accumulated Tub2p is predominantly associated with the plasma membrane.     

矩叶蓝果树的订正

据笔者对采自云南勐腊的“矩叶蓝果树”模式标本及曾被定为该变种的标本研究,该变种具聚伞花序,花梗长3－5mm,雌花子房上位,核果果皮极薄,内具1种子,并非蓝果树属植物,与茶茱萸科粗丝术属之粗丝本完全同种。粗丝木产中国西南至东南部,印度、斯里兰卡、缅甸、泰国、印度支那也有分布。     

The effect of water status and season on the incorporation of 14CO2 and [14C]-acetate into resin and rubber fractions in guayule

The effects of drought stress and season on both allocation of photosynthates to stems and leaves and potential for stem rubber synthesis were studied in guayule ( Parthenium argentatum Gray USDA line 11604). Two-year-old plants grown under field conditions in the Negev desert of Israel were subjected to different irrigation regimes, and water status was assessed by measuring the relative water content (RWC). Undetached plant tips were exposed to a 1 h pulse of ¹⁴CO₂, followed by a 24 h chase. ¹⁴C fixed and translocated to different plants parts and notably ¹⁴C incorporation into rubber and resin fractions was determined. The potential of detached branch slices to incorporate [¹⁴C]-acetate into rubber was also studied. A higher percentage of fixed ¹⁴C was translocated from shoot tips in winter (28–30%) than in summer (15–18%). The percentage of [¹⁴C]-acctate incorporated into the rubber fraction by stem slices was maximal in winter (20%) and minimal in summer (3–5%) in both cases in the absence of drought stress. In summer the translocation of photosynthates into stems was inversely related to plant RWC, dropping from 18% three days after irrigation to 3% 14 days later, and the potential of stems to synthesise rubber was high under drought conditions and low in well irrigated plants.     

NUCLEOTIDE SEQUENCE OF THE GENES FOR RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE/OXYGENASE LARGE SUBUNIT AND RIBOSOMAL PROTEIN S14 FROM A CHLORELLA-LIKE GREEN ALGA1

Two chloroplast genes were sequenced from an exsymbiotic strain of a eukaryotic, Chlorella-like green alga. The genes for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) and the ribosomal protein S14 (rps14) were oriented in the same direction and were separated by 402 bp. The rbcLs of the exsymbiont and a free living Chlorella ellipsoidea were compared with other reported rbcL sequences. The rbcL gene of the exsymbiont is closely related to that of free-living Chlorella ellipsoidea. This is the first published report of an rps 14 gene sequence from an alga.     

Involvement of endogenously produced 1,25-dihydroxyvitamin D-3 in the growth and differentiation of human keratinocytes

In this study, we investigated the possibility that cultured keratinocytes from normal human adult skin produce 1,25-dihydroxyvitamin D-3 (1,25(OH)₂D₃, a biologically active form of vitamin D-3) from 25-hydroxyvitamin D-3 [25(OH)D₃], and that 1,25(OH)₂D₃ endogenously produced by keratinocytes is involved in the self regulation of their growth and differentiation. To determine whether 1,25(OH)₂D₃ is produced from 25(OH)D₃ by skin keratinocytes, 25(OH)[³H]D₃ was added to keratinocyte cultures and incubated for 1 h and 5 h. The intracellular and extracellular metabolites were analyzed by three chromatographic systems. The three chromatograms revealed that the major metabolite produced from 25(OH)₂D₃ was 1,25(OH)₂D₃. Most of the 1,25(OH)₂D₃ endogenously produced from 25(OH)D₃ remained within the cells. To examine the time course of 1,25(OH)₂D₃ production, the amount of 1,25(OH)[³H]D₃ was measured at 15 min, 1 h, 5 h and 10 h, being at a maximum 1 h after the addition of 25(OH)D₃. These data indicate that keratinocytes rapidly convert 25(OH)D₃ to 1,25(OH)₂D₃ and that 1,25(OH)₂D₃ is not released into the medium. To determine whether endogenously produced 1,25(OH)₂D₃ is involved in the regulation of growth and differentiation of normal human keratinocytes, we examined the effects of 1,25(OH)₂D₃ and 25(OH)D₃ on their growth and differentiation. Keratinocyte growth was inhibited to 52.6% and 23.4% by 10^?8 M and 10^?7 M 1,25(OH)₂D₃ and to 80.5% and 23.9% by 10^?8 M and 10^?7 M 25(OH)D₃, respectively. Differentiation of these cells was evaluated by quantifying the number which express involucrin, a precursor protein of cornified envelope. The population of involucrin expressing cells (differentiated cells) increased from 6.2% to 14.5% by 2.5·10^?7 M 1,25(OH)₂D₃, and to 11.8% by 2.5·10^?7 M 25(OH)D₃. These results clearly indicate that 25(OH)D₃ is as effective on human keratinocytes as 1,25(OH)₂D₃ in inhibiting growth and inducing differentiation, although to a slightly lesser extent than 1,25(OH)₂D₃. The possibility that the effect of 25(OH)D₃ is mediated through binding to the 1,25(OH)₂D₃ receptor can be excluded, since a competitive binding assay revealed that the affinity of 25(OH)D₃ for the 1,25(OH)₂D₃ receptor in a cytosolic extract of keratinocytes was 100-times lower than that of 1,25(OH)₂D₃. Thus, these results suggest that 1,25(OH)₂D₃ endogenously produced in keratinocytes from 25(OH)D₃ is involved in the regulation of their growth and differentiation in vitro.     

丙烯酸对十六碳二元酸发酵的影响和16L罐扩试

本文报道丙烯酸对发酵生产十六碳二元酸(DC_(16))的影响,加入0.1%丙烯酸,DC_(16)产量提高20—30%.在16L自动控制罐上,在最佳条件下,加入20%(v/v)正十六烷(nC_(16)),发酵5天,DC_(16)高达120g/L,nC_(16)的转化率高达79%.     

PRIMARY PRODUCTION OF EDAPHIC ALGAL COMMUNITIES IN A MISSISSIPPI SALT MARSH1

Primary production rates of edaphic algae associated with the sediments beneath four monospecific canopies of vascular plants were determined over an annual cycle in a Mississippi salt marsh. The edaphic algal flora was dominated by small, motile pennate diatoms. Algal production (as measured by ¹⁴C uptake) was generally highest in spring-early summer and lowest in fall. Hourly rates ranged from a low of 1.4 mg C/m² in Juncus roemerianus Scheele to a high of 163 mg C/m² beneath the Scirpus olneyi gray canopy. Stepwise multiple regressions identified a soil moisture index and chlorophyll a as the best environmental predictors of hourly production; light energy reaching the marsh surface and sediment and air temperature proved of little value. Adding the relative abundances of 33 diatom taxa to the set of independent variables only slightly increased R²; however, virtually all variables selected were diatom taxa. R² was only 0.38 for the Spartina alterniflora Loisel. habitat but ranged from 0.70 to 0.87 for the remaining three vascular plant zones. Annual rates of algal production (g C/m²) were estimated as follows: Juncus (28), Spartina (57), Distichlis spicata (L.) Greene (88), and Scirpus (151). The ratio of annual edaphic algal production to vascular plant net aerial production (EAP / VPP) was 10–12% for the first three habitats and 61% for Scirpus. Chlorophyll a concentrations, annual algal production rates, and EAP / VPP values were comparable to those determined in Texas, Delaware, and Massachusetts salt marshes but lower than those reported for Georgia and particularly California marshes.