Biology subjects

Roth, B. L.

Publications and source records attributed to Roth, B. L..

3 recordsLinked to original sources

bioRxiv2018

A Chemogenetic Platform for Spatio-temporal Control of β-arrestin Translocation and Signaling at G protein-Coupled Receptors

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=61 SRC=\"FIGDIR/small/251769_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (15K):\norg.highwire.dtl.DTLVardef@1e4bf96org.highwire.dtl.DTLVardef@de4937org.highwire.dtl.DTLVardef@198dd33org.highwire.dtl.DTLVardef@e26f46_HPS_FORMAT_FIGEXP M_FIG C_FIG Although ligand-activated GPCRs induce both G-protein and {beta}-arrestin dependent signaling, gaining precise spatio-temporal control of {beta}-arrestin signaling has proven elusive. Here we describe a platform for specifically activating {beta}-arrestin-dependent signaling in situ. The platform, which we have dubbed \"GA-PAIR\" (GPCR/{beta}-Arrestin -Plant protein and Abscisic acid Induced Recruitment), can be controlled by the inert phytochemical S-(+)-abscisic acid (ABA). ABA induces interaction between ABI1 (ABA Insensitive 1) and PYL1 (Pyrabactin Resistance (PYR) 1-Like), two plant proteins with no mammalian counterparts. We fused ABI to the engineered human muscarinic M3 G protein-coupled receptor (hM3Dq) and PYL1 to {beta}-arrestin2. Addition of ABA induced rapid and nearly complete translocation of the PYL-{beta}-arrestin fusion protein and, importantly, induced both ERK and Akt signaling. Photo-uncaging a new photo-caged ABA analogue allowed us to gain relatively precise spatio-temporal control over {beta}-arrestin translocation. Because GA-PAIR facilitates the exclusive activation of endogenous {beta}-arrestin signaling pathways in the absence of a GPCR ligand or G protein, the GA-PAIR system will facilitate deconvoluting GPCR signaling in situ.

synthetic biology↗

bioRxiv2017

Cell-type selective deletion of RSK2 reveals insights into altered signaling in Coffin-Lowry Syndrome

Coffin-Lowry syndrome (CLS) is an X-linked syndromic form of mental retardation characterized by various skeletal dysmorphisms, moderate to severe mental retardation, and in some cases, psychosis. CLS is caused by loss-of-function mutations of the p90 ribosomal S6 kinase 2 (RPS6KA3) gene encoding a growth factor-regulated serine/threonine kinase, ribosomal S6 kinase 2 (RSK2). We previously identified RSK2 as a novel interacting protein that tonically inhibits 5-HT2A receptor signaling by phosphorylating Ser-314 within the third intracellular loop. To determine if RSK2 inhibits 5-HT2A receptor signaling in vivo and whether disruption of RSK2 could lead to schizophrenia-like behaviors - as is seen in some CLS patients - we genetically disrupted the function of RSK2 either globally or selectively in forebrain pyramidal neurons in mice. Both global and forebrain-selective RSK2 deletion augmented the locomotor responses to the psychotomimetic drugs phencyclidine (PCP) and amphetamine (AMPH). Significantly, forebrain-selective deletion of RSK2 augmented 5-HT2A receptor signaling as exemplified by enhanced 5-HT2A-mediated c-fos activation and head-twitch response without altering the levels or distribution of 5-HT2A receptor protein. Thus, RSK2 modulates 5HT2A receptor function in vivo, and disruption of RSK2 leads to augmented psychostimulant-induced responses reminiscent of those seen in many animal models of schizophrenia. These findings strengthen the association between 5-HT2A receptor dysfunction and psychosis, and provide a potential mechanism underlying the schizophrenia-like symptoms present in some CLS patients.\n\nHighlightsO_LIGlobal and cell-type-specific RSK2 knock-out mice were assessed behaviorally and pharmacologically\nC_LIO_LIAugmentation of amphetamine and PCP locomotor responses were seen in both global and forebrain-specific RSK2 KO mice\nC_LIO_LIAugmentation of 5-HT2A serotonin receptor function but not number was also observed\nC_LIO_LIThese alterations reveal insights into mechanisms potentially responsible for behavioral sequlae of Coffin-Lowry Syndrome\nC_LI

neuroscience↗

bioRxiv2017

A Simple Representation Of Three-Dimensional Molecular Structure

Statistical and machine learning approaches predict drug-to-target relationships from 2D small-molecule topology patterns. One might expect 3D information to improve these calculations. Here we apply the logic of the Extended Connectivity FingerPrint (ECFP) to develop a rapid, alignment-invariant 3D representation of molecular conformers, the Extended Three-Dimensional FingerPrint (E3FP). By integrating E3FP with the Similarity Ensemble Approach (SEA), we achieve higher precision-recall performance relative to SEA with ECFP on ChEMBL20, and equivalent receiver operating characteristic performance. We identify classes of molecules for which E3FP is a better predictor of similarity in bioactivity than is ECFP. Finally, we report novel drug-to-target binding predictions inaccessible by 2D fingerprints and confirm three of them experimentally with ligand efficiencies from 0.442 - 0.637 kcal/mol/heavy atom.

bioinformatics↗