Papers by David Siderovski
Journal of Biological Chemistry, Nov 1, 2014
Background: NLRP3 is a key regulator of innate inflammation and is linked to inflammatory disease... more Background: NLRP3 is a key regulator of innate inflammation and is linked to inflammatory diseases. Results: GPSM3 associates with NLRP3 and inhibits its function. Conclusion: GPSM3 specifically inhibits NLRP3-dependent inflammasome activity by interacting with its leucine-rich repeat domain. Significance: This association uncovers a putative new mechanism of NLRP3 control, linking a G protein modulator to NLRP3-dependent inflammatory diseases. Inflammasomes are multi-protein complexes that regulate maturation of the interleukin 1-related cytokines IL-1 and IL-18 through activation of the cysteine proteinase caspase-1. NOD-like receptor family, pyrin domain containing 3 (NLRP3) protein is a key component of inflammasomes that assemble in response to a wide variety of endogenous and pathogen-derived danger signals. Activation of the NLRP3-inflammasome and subsequent secretion of IL-1 is highly regulated by at least three processes: transcriptional activation of both NLRP3 and pro-IL-1 genes, non-transcriptional priming of NLRP3, and final activation of NLRP3. NLRP3 is predominantly expressed in cells of the hematopoietic lineage. Using a yeast two-hybrid screen, we identified the hematopoietic-restricted protein, G protein signaling modulator-3 (GPSM3), as a NLRP3-interacting protein and a negative regulator of IL-1 production triggered by NLRP3-dependent inflammasome activators. In monocytes, GPSM3 associates with the C-terminal leucine-rich repeat domain of NLRP3. Bone marrow-derived macrophages lacking GPSM3 expression exhibit an increase in NLRP3-dependent IL-1, but not TNF-␣, secretion. Furthermore, GPSM3-null mice have enhanced serum and peritoneal IL-1 production following Alum-induced peritonitis. Our findings suggest that GPSM3 acts as a direct negative regulator of NLRP3 function. * This work was supported by National Institutes of Health Grants R01 AI088255 (to J. A. D.), U19 AI109965 (to J. A. D. and J. P. T.), and U54 GM104942 (to D. P. S.) via the WVCTSI Pilot Grants Program and by a Burroughs Wellcome Fund Career Award for Medical Scientists (to J. A. D.). † We are grateful to Dr. Chahnaz Kebaier (deceased, formerly of the University of North Carolina at Chapel Hill) for the work she performed studying bone marrow-derived macrophages for these studies.
Recent Advances in Therapeutic Drug Monitoring and Clinical Toxicology
We describe the isolation of human LH-2, a putative transcription factor containing two cysteine-... more We describe the isolation of human LH-2, a putative transcription factor containing two cysteine-rich regions (LIM domains) and a homeobox (Hox) DNA-binding domain. High levels of hLH-2 expression were observed in all cases of chronic myelogenous leukaemia (CML) tested, regardless of disease status. hLH-2 was mapped to chromosome 9Q33-34.1, in the same region as the reciprocal translocation that creates the BCR-ABL chimera of the Philadelphia chromosome (Ph'), the hallmark of CML; hLH-2 was retained on the derivative 9 chromosome and is therefore centromeric of c-ABL. The proximity of hLH-2 to the breakpoint on chromosome 9 raises the possibility of cis-activation by the t(9;22)(q34;q11) translocation. In addition to finding hLH-2 expression in all cases of CML, expression was observed in lymphoid malignancies and myeloid cell lines, but not in primary cases of acute myelogenous leukaemia. The role of hLH-2 in the development or progression of leukaemia is not known. However, hL...
PLC-epsilon was identified recently as a phosphoinositide-hydrolyzing phospholipase C (PLC) conta... more PLC-epsilon was identified recently as a phosphoinositide-hydrolyzing phospholipase C (PLC) containing catalytic domains (X, Y, and C2) common to all PLC isozymes as well as unique CDC25- and Ras-associating domains. Novel regulation of this PLC isozyme by the Ras oncoprotein and alpha-subunits (Galpha(12)) of heterotrimeric G proteins was illustrated. Sequence analyses of PLC-epsilon revealed previously unrecognized PH and EF-hand domains in the amino terminus. The known interaction of Gbetagamma subunits with the PH domains of other proteins led us to examine the capacity of Gbetagamma to activate PLC-epsilon. Co-expression of Gbeta(1)gamma(2) with PLC-epsilon in COS-7 cells resulted in marked stimulation of phospholipase C activity. Gbeta(2) and Gbeta(4) in combination with Ggamma(1), Ggamma(2), Ggamma(3), or Ggamma(13) also activated PLC-epsilon to levels similar to those observed with Gbeta(1)-containing dimers of these Ggamma-subunits. Gbeta(3) in combination with the same Gga...
A basic helix-loop-helix phosphoprotein gene, G0S8, was recently isolated by differential screeni... more A basic helix-loop-helix phosphoprotein gene, G0S8, was recently isolated by differential screening of cDNA from human blood mononuclear cells stimulated with a T cell mitogen and cycloheximide. In this study, G0S8 expression was examined in normal and malignant hematopoietic cells by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). G0S8 expression was observed in most fresh samples of acute myelogenous leukemia (AML) (28/30) and most cases of adult acute lymphoblastic leukemia (ALL) (9/11) regardless of clinical classification. G0S8 mRNA was also detected in all cases tested of chronic myelogenous leukemia (CML) in blast crisis. However, G0S8 expression was not detected in CML patients in chronic phase, nor in normal bone marrow or other hematopoietic cells. G0S8 has been mapped using fluorescence in situ hybridization (FISH) to human chromosome 1q31, the same site reported for the B cell homolog BL34/1R20 and within a region implicated in the de...
The Journal of biological chemistry, 2001
Mutation of Galpha(q) or Galpha(s) N-terminal contact sites for Gbetagamma resulted in alpha subu... more Mutation of Galpha(q) or Galpha(s) N-terminal contact sites for Gbetagamma resulted in alpha subunits that failed to localize at the plasma membrane or undergo palmitoylation when expressed in HEK293 cells. We now show that overexpression of specific betagamma subunits can recover plasma membrane localization and palmitoylation of the betagamma-binding-deficient mutants of alpha(s) or alpha(q). Thus, the betagamma-binding-defective alpha is completely dependent on co-expression of exogenous betagamma for proper membrane localization. In this report, we examined the ability of beta(1-5) in combination with gamma(2) or gamma(3) to promote proper localization and palmitoylation of mutant alpha(s) or alpha(q). Immunofluorescence localization, cellular fractionation, and palmitate labeling revealed distinct subtype-specific differences in betagamma interactions with alpha subunits. These studies demonstrate that 1) alpha and betagamma reciprocally promote the plasma membrane targeting of...
Genes and immunity, 2016
G protein signaling modulator 3 (GPSM3) is a regulator of G protein-coupled receptor signaling, w... more G protein signaling modulator 3 (GPSM3) is a regulator of G protein-coupled receptor signaling, with expression restricted to leukocytes and lymphoid organs. Previous genome-wide association studies have highlighted single-nucleotide polymorphisms (SNPs; rs204989 and rs204991) in a region upstream of the GPSM3 transcription start site as being inversely correlated to the prevalence of rheumatoid arthritis (RA)-this association is supported by the protection afforded to Gpsm3-deficient mice in models of inflammatory arthritis. Here, we assessed the functional consequences of these polymorphisms. We collected biospecimens from 50 volunteers with RA diagnoses, 50 RA-free volunteers matched to the aforementioned group and 100 unmatched healthy young volunteers. We genotyped these individuals for GPSM3 (rs204989, rs204991), CCL21 (rs2812378) and HLA gene region (rs6457620) polymorphisms, and found no significant differences in minor allele frequencies between the RA and disease-free coho...
Nature, 1999
Mitochondria play a key part in the regulation of apoptosis (cell death) 1, 2 . Their intermembra... more Mitochondria play a key part in the regulation of apoptosis (cell death) 1, 2 . Their intermembrane space contains several proteins that are liberated through the outer membrane in order to participate in the degradation phase of apoptosis 3, 4, 5, 6, 7, 8, 9 . Here we report the identification ...
Current Biology, 1996
Organisms as diverse as fungi and humans use G-protein-coupled receptors to control signal transd... more Organisms as diverse as fungi and humans use G-protein-coupled receptors to control signal transduction pathways responsive to various hormones, neuroregulatory molecules and other sensory stimuli [1]. Continual stimulation of these receptors often leads to their ...
Molecular and Biochemical Parasitology, 1993
Molecular and Biochemical Parasitology, 1993
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Papers by David Siderovski