eEF-2 Phosphorylation Down-Regulates P-Glycoprotein Over-Expression in Rat Brain Microvessel Endothelial Cells

ObjectiveWe investigated whether glutamate, NMDA receptors, and eukaryote elongation factor-2 kinase (eEF-2K)/eEF-2 regulate P-glycoprotein expression, and the effects of the eEF-2K inhibitor NH125 on the expression of P-glycoprotein in rat brain microvessel endothelial cells (RBMECs).MethodsCortex was obtained from newborn Wistar rat brains. After surface vessels and meninges were removed, the pellet containing microvessels was resuspended and incubated at 37°C in culture medium. Cell viability was assessed by the MTT assay. RBMECs were identified by immunohistochemistry with anti-vWF. P-glycoprotein, phospho-eEF-2, and eEF-2 expression were determined by western blot analysis. Mdr1a gene expression was analyzed by RT-PCR.ResultsMdr1a mRNA, P-glycoprotein and phospho-eEF-2 expression increased in L-glutamate stimulated RBMECs. P-glycoprotein and phospho-eEF-2 expression were down-regulated after NH125 treatment in L-glutamate stimulated RBMECs.ConclusionseEF-2K/eEF-2 should have played an important role in the regulation of P-glycoprotein expression in RBMECs. eEF-2K inhibitor NH125 could serve as an efficacious anti-multidrug resistant agent.     

Mutational analysis of slyD, an Escherichia coli gene encoding a protein of the FKBP immunophilin family

slyD encodes a 196 amino acid polypeptide that is a member of the FKBP family of cis–trans peptidyl–prolyl isomerases (PPIases). slyD mutations affect plaque formation by the phage φX174 by blocking the action of the phage lysis protein E. Here we describe the selection of a set of spontaneous slyD mutations conferring resistance to the expression of gene E from a plasmid. These mutations occur disproportionately in residues of SlyD that, based on the structure of the prototype mammalian FKBP12, make ligand contacts with immunosuppressing drug molecules or are conserved in other FKBP proteins. A wide variation in the plating efficiency of φX174 on these E  ^R strains is observed, relative to the parental, indicating that these alleles differ widely in residual SlyD activity. Moreover, it is found that slyD mutations cause significant growth rate defects in Escherichia coli B and C backgrounds. Finally, overexpression of slyD causes filamentation of the host. Thus, among the FKBP genes found in organisms across the evolutionary spectrum, slyD is unique in having three distinct drug-independent phenotypes.     

Mapping QTLs and meta-QTLs for two inflorescence architecture traits in multiple maize populations under different watering environments

Drought significantly affects the architectural development of maize inflorescence, which leads to massive losses in grain yield. However, the genetic mechanism for traits involved in inflorescence architecture in different watering environments, remains poorly understood in maize. In this study, 19 QTLs for tassel primary branch number (TBN) and ear number per plant (EN) were detected in 2 F_2:3 populations under both well-watered and water-stressed environments by single environment mapping with composite interval mapping (CIM); 11/19 QTLs were detected under water-stressed environments. Moreover, 21 QTLs were identified in the 2 F_2:3 populations by joint analysis of all environments with a mixed linear model based on composite interval mapping (MCIM), 11 QTLs were involved in QTL × environment interactions, seven epistatic interactions were identified with additive by additive/dominance effects. Remarkably, 12 stable QTLs (sQTLs) were simultaneously detected by single environment mapping with CIM and joint analysis through MCIM, which were concentrated in ten bins across the chromosomes: 1.05_1.07, 1.08_1.10, 2.01_2.04, 3.01, 4.06, 4.09, 5.06_5.07, 6.05, 7.00, and 7.04 regions. Twenty meta-QTLs (mQTLs) were detected across 19 populations under 51 watering environments using a meta-analysis, and 34 candidate genes were predicted in corresponding mQTLs regions to be involved in the regulation of inflorescence development and drought resistance. Therefore, these results provide valuable information for finding quantitative trait genes and to reveal the genetic mechanisms responsible for TBN and EN under different watering environments. Furthermore, alleles for TBN and EN provide useful targets for marker-assisted selection to generate high-yielding maize varieties.     

A Modified BFGS Formula Using a Trust Region Model for Nonsmooth Convex Minimizations

This paper proposes a modified BFGS formula using a trust region model for solving nonsmooth convex minimizations by using the Moreau-Yosida regularization (smoothing) approach and a new secant equation with a BFGS update formula. Our algorithm uses the function value information and gradient value information to compute the Hessian. The Hessian matrix is updated by the BFGS formula rather than using second-order information of the function, thus decreasing the workload and time involved in the computation. Under suitable conditions, the algorithm converges globally to an optimal solution. Numerical results show that this algorithm can successfully solve nonsmooth unconstrained convex problems.