Helix 11 Dynamics Is Critical for Constitutive Androstane Receptor Activity

SummaryThe constitutive androstane receptor (CAR) transactivation can occur in the absence of exogenous ligand and this activity is enhanced by agonists TCPOBOP and meclizine. We use biophysical and cell-based assays to show that increased activity of CAR(TCPOBOP) relative to CAR(meclizine) corresponds to a higher affinity of CAR(TCPOBOP) for the steroid receptor coactivator-1. Additionally, steady-state fluorescence spectra suggest conformational differences between CAR(TCPOBOP):RXR and CAR(meclizine):RXR. Hydrogen/deuterium exchange (HDX) data indicate that the CAR activation function 2 (AF-2) is more stable in CAR(TCPOBOP):RXR and CAR(meclizine):RXR than in CAR:RXR. HDX kinetics also show significant differences between CAR(TCPOBOP):RXR and CAR(meclizine):RXR. Unlike CAR(meclizine):RXR, CAR(TCPOBOP):RXR shows a higher overall stabilization that extends into RXR. We identify residues 339–345 in CAR as an allosteric regulatory site with a greater magnitude reduction in exchange kinetics in CAR(TCPOBOP):RXR than CAR(meclizine):RXR. Accordingly, assays with mutations on CAR at leucine-340 and leucine-343 confirm this region as an important determinant of CAR activity

The Stern diatomic sequence via generalized Chebyshev polynomials

Let a(n) be the Stern's diatomic sequence, and let x1,...,xr be the distances between successive 1's in the binary expansion of the (odd) positive integer n. We show that a(n) is obtained by evaluating generalized Chebyshev polynomials when the variables are given the values x1+1, ..., xr+1, and we derive a formula expressing the same polynomials in terms of sets of increasing integers of alternating parity. We also show that a(n) = Det(Ir + Mr), where Ir is the rxr identity matrix, and Mr is the rxr matrix that has x1,...,xr along the main diagonal, then all 1's just above and below the main diagonal, and all the other entries are 0

Magreceptorok működésének genomszintü vizsgálata kromatin immunprecipitációval primer humán immun sejtekben = Decoding nuclear hormone receptor activity using chromatin immunoprecipitation in human primary immune cells

A kutatómunka során szisztematikusan feltérképeztük az un RXR heterodimer típusú receptorok szerepét a humán monocita eredetű dendritikus sejtek differenciálódása és immunfunkciói során. Ezek a receptorok a PPARg, LXR, VDR és RAR receptorok volta, melyeket kiegészítettünk az RXR receptorral is. A specikus utvonalak azonosítása mellett általános megállapításként elmondhattuk, hogy ez a receptorcsalad olyan molekuláris szenzorként működik, mely képes átprogramozni a sejt genkifejeződését külső és belső lipid szintek változása során. Összefüggést talaltunk az IL4 STAT6-on kerestüli szignalizációja és a PPARg receptor között és legújabban feltérképeztük a az RXR receptor genomi kötőhelyeit ls cisztromikus kölcsönhatásait is. | During our work we have systematically mapped the so called RXR heterodimeric receptors roles in monocyte derived dendritic cells during differentiation and immune function. These receptors included PPARg, LXR, VDR and RXR and also included their heterodimeric partner RXR. Based on the data obtained one can conclude that besides the specialized pathways identified, these receptors act as molecular sensors in detecting changing extra and intracellular lipid levels. We have also identified an interaction between IL4 mediated STAT6 signaling and PPARg and most recently we have detrained the RXR cistrome and its interactions

A Pleiotropic Response Is Induced in F9 Embryonal Carcinoma Cells and Rhino Mouse Skin by All-trans-Retinoic Acid, a RAR Agonist but Not by SR11237, a RXR-Selective Agonist

We evaluated SR11237, a retinoid X receptor (RXR)- specific compound, for its pharmacologic effects on cell differentiation in F9 embryonal carcinoma cells and rhino mouse epidermis. SR11237 can cause RXR/RXR homodimers to form and transactivate a reporter gene containing a RXR-response element. We confirmed, using nuclear receptor co-transfection assays in COS-1 cells, that SR11237 is effective at transactivating a chloramphenicol acetyltransferase reporter gene through RXRs but not retinoic acid receptors. When SR11237 was tested for its ability to modulate cell differentiation, it was inactive on F9 embryonal carcinoma cells and rhino mouse skin. Because differentiation in these systems is known to be regulated by RAR-specific compounds, such as all-trans-retinoic acid and (E)-4-[2- (5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1- propenyl benzoic acid], our results with SR11237 are compatible with the concept that classical retinoid pleiotropic responses are mediated by RXR/RAR heterodimeric nuclear receptors rather than through RXR/RXR homodimers

Finite sample properties of multiple imputation estimators

Finite sample properties of multiple imputation estimators under the linear regression model are studied. The exact bias of the multiple imputation variance estimator is presented. A method of reducing the bias is presented and simulation is used to make comparisons. We also show that the suggested method can be used for a general class of linear estimators

Cross-Talk between PPARs and the Partners of RXR: A Molecular Perspective

The PPARs are integral parts of the RXR-dependent signaling networks. Many other nuclear receptor subfamily 1 members also require RXR as their obligatory heterodimerization partner and they are often co-expressed in any given tissue. Therefore, the PPARs often complete with other RXR-dependent nuclear receptors and this competition has important biological implications. Thorough understanding of this cross-talk at the molecular level is crucial to determine the detailed functional roles of the PPARs. At the level of DNA binding, most RXR heterodimers bind selectively to the well-known “DR1 to 5” DNA response elements. As a result, many heterodimers share the same DR element and must complete with each other for DNA binding. At the level of heterodimerization, the partners of RXR share the same RXR dimerization interface. As a result, individual nuclear receptors must complete with each other for RXR to form functional heterodimers. Cross-talk through DNA binding and RXR heterodimerization present challenges to the study of these nuclear receptors that cannot be adequately addressed by current experimental approaches. Novel tools, such as engineered nuclear receptors with altered dimerization properties, are currently being developed. These tools will enable future studies to dissect specific RXR heterodimers and their signaling pathways

Beyond the Min-Cut Bound: Deterministic Network Coding for Asynchronous Multirate Broadcast

In a single hop broadcast packet erasure network, we demonstrate that it is possible to provide multirate packet delivery outside of what is given by the network min-cut. This is achieved by using a deterministic non-block-based network coding scheme, which allows us to sidestep some of the limitations put in place by the block coding model used to determine the network capacity. Under the network coding scheme we outline, the sender is able to transmit network coded packets above the channel rate of some receivers, while ensuring that they still experience nonzero delivery rates. Interestingly, in this generalised form of asynchronous network coded broadcast, receivers are not required to obtain knowledge of all packets transmitted so far. Instead, causal feedback from the receivers about packet erasures is used by the sender to determine a network coded transmission that will allow at least one, but often multiple receivers, to deliver their next needed packet. Although the analysis of deterministic coding schemes is generally a difficult problem, by making some approximations we are able to obtain tractable estimates of the receivers' delivery rates, which are shown to match reasonably well with simulation. Using these estimates, we design a fairness algorithm that allocates the sender's resources so all receivers will experience fair delivery rate performance

A review about lycopene-induced nuclear hormone receptor signalling in inflammation and lipid metabolism via still unknown endogenous apo-10´-lycopenoids

Lycopene is the red pigment in tomatoes and tomato products and is an important dietary carotenoid found in the human organism. Lycopene-isomers, oxidative lycopene metabolites and apo-lycopenoids are found in the food matrix. Lycopene intake derived from tomato consumption is associated with alteration of lipid metabolism and a lower incidence of cardiovascular diseases (CVD). Lycopene is mainly described as a potent antioxidant but novel studies are shifting towards its metabolites and their capacity to mediate nuclear receptor signalling. Di-/tetra-hydro-derivatives of apo-10´-lycopenoic acid and apo-15´-lycopenoic acids are potential novel endogenous mammalian lycopene metabolites which may act as ligands for nuclear hormone mediated activation and signalling. In this review, we postulate that complex lycopene metabolism results in various lycopene metabolites which have the ability to mediate transactivation of various nuclear hormone receptors like RARs, RXRs and PPARs. A new mechanistic explanation of how tomato consumption could positively modulate inflammation and lipid metabolism is discussed

The anti-cancer effect of retinoic acid signaling in CRC occurs via decreased growth of ALDH+ colon cancer stem cells and increased differentiation of stem cells

Background: Tumorigenesis is driven by stem cell (SC) overpopulation. BecauseALDH is both a marker for SCs in many tissues and a key enzyme in retinoid acid (RA)signaling, we studied RA signaling in normal and malignant colonic SCs.Hypothesis: RA signaling regulates growth and differentiation of ALDH+ colonicSCs dysregulation of RA signaling contributes to SC overpopulation and colorectalcancer (CRC) development.Methods: We analyzed normal and malignant colonic tissues and CRC cell linesto see if retinoid receptors (RXR &RAR) are exclusively expressed in ALDH+ SCs,and if RA signaling changes during CRC development. We determined whether RAsignaling regulates cancer SC (CSC) proliferation, differentiation, sphere formation,and population size.Results: RXR &RAR were expressed in ALDH+ colonic SCs, but not in MCM2+proliferative cells. Western blotting/immunostaining of CRCs revealed that RAsignaling components become overexpressed in parallel with ALDH overexpression,which coincides with the known overpopulation of ALDH+ SCs that occurs during,and drives, CRC development. Treatment of SCs with all-trans retinoic acid (ATRA)decreased proliferation, sphere formation and ALDH+ SC population size, and induceddifferentiation along the neuroendocrine cell (NEC) lineage.Conclusions: Retinoid signaling, by regulating ALDH+ colonic CSCs, decreases SCproliferation, sphere formation, and population size, and increases SC differentiation toNECs. Dysregulation of RA signaling in colonic SCs likely contributes to overpopulationof ALDH+ SCs and CRC growth.Implications: That retinoid receptors RXR and RAR are selectively expressed inALDH+ SCs indicates RA signaling mainly occurs via ALDH+ SCs, which provides amechanism to selectively target CSCs. © 2018 Impact Journals LLC. All rights reserved

Structure-based analysis of the ultraspiracle protein and docking studies of putative ligands

The ultraspiracle protein (USP) is the insect ortholog of the mammalian retinoid X receptor (RXR). Fundamental questions concern the functional role of USP as the heterodimerization partner of insect nuclear receptors such as the ecdysone receptor. The crystallographic structures of the ligand binding domain of USPs of Heliothis virescens and Drosophila melanogaster solved recently show that helix 12 is locked in an antagonist conformation raising the question whether USPs could adopt an agonist conformation as observed in RXRα. In order to investigate this hypothesis, a homology model for USP is proposed that allows a structural analysis of the agonist conformation of helix 12 based on the sequence comparison with RXR. For USP, one of the main issues concerns its function and in particular whether its activity is ligand independent or not. The x-ray structures strongly suggest that USP can bind ligands. Putative ligands have therefore been docked in the USP homology model. Juvenile hormones and juvenile hormone analogs were chosen as target ligands for the docking study. The interaction between the ligand and the receptor are examined in terms of the pocket shape as well as in terms of the chemical nature of the residues lining the ligand binding cavity