Enhanced citric acid production by a yeast <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> over-expressing a pyruvate carboxylase gene

In this study, after the expression of a pyruvate carboxylase gene (PYC) cloned from Meyerozyma guilliermondii in a marine-derived yeast Yarrowia lipolytica SWJ-1b, a transformant PG86 obtained had much higher PYC activity than Y. lipolytica SWJ-1b. At the same time, the PYC gene expression and citric acid (CA) production by the transformant PG86 were also greatly enhanced. When glucose concentration in the medium was 60.0 g L^?1, CA concentration formed by the transformant PG86 was 34.02 g L^?1, leading to a CA yield of 0.57 g g^?1 of glucose. During a 10-L fed-batch fermentation, the final concentration of CA was 101.0 ± 1.3 g L^?1, the yield was 0.89 g g^?1 of glucose, the productivity was 0.42 g L^?1 h^?1 and only 5.93 g L^?1 reducing sugar was left in the fermented medium within 240 h of the fed-batch fermentation. HPLC analysis showed that most of the fermentation products were CA.     

Cinderella story: PI4P goes from precursor to key signaling molecule

Abstract

Phosphatidylinositol lipids are signaling molecules involved in nearly all aspects of cellular regulation. Production of phosphatidylinositol 4-phosphate (PI4P) has long been recognized as one of the first steps in generating poly-phosphatidylinositol phosphates involved in actin organization, cell migration, and signal transduction. In addition, progress over the last decade has brought to light independent roles for PI4P in membrane trafficking and lipid homeostasis. Here, we describe recent advances that reveal the breadth of processes regulated by PI4P, the spectrum of PI4P effectors, and the mechanisms of spatiotemporal control that coordinate crosstalk between PI4P and cellular signaling pathways.     

Phosphorylation of Pkp1 by RIPK4 regulates epidermal differentiation and skin tumorigenesis

Tissue homeostasis of skin is sustained by epidermal progenitor cells localized within the basal layer of the skin epithelium. Post‐translational modification of the proteome, such as protein phosphorylation, plays a fundamental role in the regulation of stemness and differentiation of somatic stem cells. However, it remains unclear how phosphoproteomic changes occur and contribute to epidermal differentiation. In this study, we survey the epidermal cell differentiation in a systematic manner by combining quantitative phosphoproteomics with mammalian kinome cDNA library screen. This approach identified a key signaling event, phosphorylation of a desmosome component, PKP1 (plakophilin‐1) by RIPK4 (receptor‐interacting serine–threonine kinase 4) during epidermal differentiation. With genome‐editing and mouse genetics approach, we show that loss of function of either Pkp1 or Ripk4 impairs skin differentiation and enhances epidermal carcinogenesis in vivo. Phosphorylation of PKP1's N‐terminal domain by RIPK4 is essential for their role in epidermal differentiation. Taken together, our study presents a global view of phosphoproteomic changes that occur during epidermal differentiation, and identifies RIPK‐PKP1 signaling as novel axis involved in skin stratification and tumorigenesis.     

Source-to-sink relationship between green leaves and green pseudobulbs of C<Subscript>3</Subscript> orchid in regulation of photosynthesis

In this paper, photosynthetic characteristics of green leaves (GL) and green pseudobulbs (GPSB) of C₃ orchid Oncidium Golden Wish were first studied. Light saturation for photosynthesis and maximum photosynthetic rates (P _max) were significantly higher in GL than in GPSB. The results of the optimal PSII quantum yield (F_v/F_m ratio), electron transport rate (ETR), the effective photochemical quantum yield (ΔF/F_m′) and nonphotochemical quenching (NPQ) of Chl fluorescence revealed that GPSB had lower light utilization than that of GL. Significantly higher photosynthetic pigments were found in GL than in GPSB. Alteration of source/sink ratio had no impact on all photosynthetic parameters for both GL and GPSB after a short term of 3 days or even a long term of 2 weeks of treatments although there were significant decreases in GL carbohydrate concentration of GL-darkened plants by the end of the day. However, decreases of all photosynthetic parameters of GL were observed in GL-darkened plants after 4 weeks of treatment compared to those of fully illuminated (FI) and GPSB-darkened plants. These results indicate that the level of carbohydrates in GL plays an important role in regulating their photosynthesis. Due to their lower photosynthetic capacities, GPSB function mainly as sinks. Darkening GPSB up to 2 weeks did not affect their own P _max and the P _max of GL and thus, did not result in significant decreases of total carbohydrate concentration of GPSB. As GPSB store a large amount of carbohydrates, it could also act as a source when the level of carbohydrates decreased. Thus, GL could depend on GPSB carbohydrates to regulate their photosynthesis when their source capacity was removed. However, 4 weeks after treatments, photosynthetic capacities of GL were significantly lower in GL- and GPSB-darkened plants than in FI plants, which could be due to the lower total soluble and insoluble sugar concentrations of both GL and GPSB in these plants.     

Hypoxia induces beta-amyloid in association with death of RGC-5 cells in culture

Beta-amyloid (Aβ) derived from amyloid precursor protein (APP) has been associated with retinal degeneration in Alzheimer’s disease (AD) and glaucoma. This study examined whether hypoxia exposure induces Aβ accumulation in RGC-5 cells. While levels of APP mRNA and protein significantly increased in the cells, elevated abundance of Aβ was also observed in cells and culture medium between 12 or 24 and 48 h after 5% O₂ hypoxia treatment. Additionally, there is a close relationship between induction of APP and Aβ and intracellular accumulation of ROS along with loss of mitochondrial membrane potential followed by the death of RGC-5 cells in culture under hypoxia. These results suggest a possible involvement of APP and Aβ in the death of RGCs challenged by hypoxia.     

分子标记育种在我国的研究进展

现代生物技术的发展为利用外源基因培育优良作物品种提供了新的途径。简要综述了分子标记育种在我国研究的总体情况以及我国在分子标记育种领域取得的成就,并对分子标记育种的前景进行了展望。     

Activation of ROCK by RhoA is regulated by cell adhesion, shape, and cytoskeletal tension

Adhesion to the extracellular matrix regulates numerous changes in the actin cytoskeleton by regulating the activity of the Rho family of small GTPases. Here, we report that adhesion and the associated changes in cell shape and cytoskeletal tension are all required for GTP-bound RhoA to activate its downstream effector, ROCK. Using an in vitro kinase assay for endogenous ROCK, we found that cells in suspension, attached on substrates coated with low density fibronectin, or on spreading-restrictive micropatterned islands all exhibited low ROCK activity and correspondingly low myosin light chain phosphorylation, in the face of high levels of GTP-bound RhoA. In contrast, allowing cells to spread against substrates rescued ROCK and myosin activity. Interestingly, inhibition of tension with cytochalasin D or blebbistatin also inhibited ROCK activity within 20 min. The abrogation of ROCK activity by cell detachment or inhibition of tension could not be rescued by constitutively active RhoA-V14. These results suggest the existence of a feedback loop between cytoskeletal tension, adhesion maturation, and ROCK signaling that likely contributes to numerous mechanochemical processes.