Characterization of desnutrin functional domains: critical residues for triacylglycerol hydrolysis in cultured cells

Murine desnutrin/human ATGL is a triacylglycerol (TAG) hydrolase with a predicted catalytic dyad within an α-β hydrolase fold in the N-terminal region. In humans, mutations resulting in C-terminal truncation cause neutral lipid storage disease with myopathy. To identify critical functional domains, we measured TAG breakdown in cultured cells by mutated or truncated desnutrin. In vitro, C-terminally truncated desnutrin displayed an even higher apparent V_max than the full-length form without changes in K_m, which may be explained by our finding of an interaction between the C- and N-terminal domains. In live cells, however, C-terminally truncated adenoviral desnutrin had lower TAG hydrolase activity. We investigated a role for the phosphorylation of C-terminal S406 and S430 residues but found that these were not necessary for TAG breakdown or lipid droplet localization in cells. The predicted N-terminal active sites, S47 and D166, were both critical for TAG hydrolysis in live cells and in vitro. We also identified two overlapping N-terminal motifs that predict lipid substrate binding domains, a glycine-rich motif (underlined) and an amphipathic α-helix (bold) within amino acid residues 10–24 (ISFAGCGFLGVYHIG). G14, F17, L18, and V20, but not G16 and G19, were important for TAG hydrolysis, suggesting a potential role for the amphipathic α-helix in TAG binding. This study identifies for the first time critical sites in the N-terminal region of desnutrin and reveals the requirement of the C-terminal region for TAG hydrolysis in cultured cells.     

Tyrosine Phosphorylation of the p21 Cyclin-dependent Kinase Inhibitor Facilitates the Development of Proneural Glioma

Phosphorylation of Tyr-88/Tyr-89 in the 3₁₀ helix of p27 reduces its cyclin-dependent kinase (CDK) inhibitory activity. This modification does not affect the interaction of p27 with cyclin-CDK complexes but does interfere with van der Waals and hydrogen bond contacts between p27 and amino acids in the catalytic cleft of the CDK. Thus, it had been suggested that phosphorylation of this site could switch the tumor-suppressive CDK inhibitory activity to an oncogenic activity. Here, we examined this hypothesis in the RCAS-PDGF-HA/nestin-TvA proneural glioma mouse model, in which p21 facilitates accumulation of nuclear cyclin D1-CDK4 and promotes tumor development. In these tumor cells, approximately one-third of the p21 is phosphorylated at Tyr-76 in the 3₁₀ helix. Mutation of this residue to glutamate reduced inhibitory activity in vitro. Mutation of this residue to phenylalanine reduced the tumor-promoting activity of p21 in the animal model, whereas glutamate or alanine substitution allowed tumor formation. Consequently, we conclude that tyrosine phosphorylation contributes to the conversion of CDK inhibitors from tumor-suppressive roles to oncogenic roles.     

Community structure and associated networks of endophytic bacteria in pea roots throughout plant life cycle

Plant and Soil - This study aimed to explore the phytoremediation potential of Salvia tiliifolia for cadmium (Cd)-contaminated soils and to understand the interactions in soil-Cd-plant systems. We...     

常绿阔叶林外生、内生菌根树种细根化学计量学性状对N添加的响应

为揭示不同菌根类型树种细根化学计量学性状对N添加的塑性响应,在福建省建瓯市万木林自然保护区常绿阔叶林内选择外生菌根树种罗浮栲(Castanopsis faberi)和内生菌根树种木荷(Schima superba)为研究对象,采用根袋法开展N添加试验,细根在根袋内生长半年后测定化学计量学指标(C、N、P、C/N、N/P、C/P)。结果表明:根序对细根化学计量学性状有显著影响,随着根序的增加,罗浮栲与木荷细根C浓度、C/N、C/P明显增加,N浓度与P浓度明显下降。N添加对细根C、N浓度均有极显著的促进作用,但对细根P浓度影响不显著,从而导致细根C/N维持稳定,但N/P、C/P升高,细根受P限制增加。细根化学计量学性状对N添加的塑性响应在不同序级间以及在外生菌根树种罗浮栲和内生菌根树种木荷之间均无显著差异。结论表明,研究所选内生、外生菌根树种细根化学计量学性状对N添加具有基本相似的响应。     

Cytosolic cleaved PINK1 represses Parkin translocation to mitochondria and mitophagy

PINK1 is a mitochondrial kinase proposed to have a role in the pathogenesis of Parkinson''s disease through the regulation of mitophagy. Here, we show that the PINK1 main cleavage product, PINK1_52, after being generated inside mitochondria, can exit these organelles and localize to the cytosol, where it is not only destined for degradation by the proteasome but binds to Parkin. The interaction of cytosolic PINK1 with Parkin represses Parkin translocation to the mitochondria and subsequent mitophagy. Our work therefore highlights the existence of two cellular pools of PINK1 that have different effects on Parkin translocation and mitophagy.     

Analysis of the bacterial community in a full-scale printing and dyeing wastewater treatment system based on T-RFLP and 454 pyrosequencing

In this study, the bacterial dynamics and structure compositions in the two-stage biological process of a full-scale printing and dyeing wastewater (PDW) treatment system were traced and analyzed by terminal restriction fragment length polymorphism (T-RFLP) and 454 pyrosequencing techniques. T-RFLP analysis showed that the microbial communities experienced significant variation in the process of seed sludge adaptation to the PDW environments and were in constant evolution during the whole running period of the system, despite the constant COD and color removal effects. Pyrosequencing results indicated that the two-stage biological system harbored rather diverse bacteria, with Proteobacteria being the predominant phylum during the steady running period, although its microbial compositions differed. The first-stage aerobic tank was dominated by α-Proteobacteria (89.05% of Proteobacteria), whereas in the second-stage aerobic tank, β- and γ-Proteobacteria, besides α-Proteobacteria, were the dominant bacterial populations.