BACKGROUND: Reactive oxygen species, such as superoxide (O(2)(-)), are involved in the abnormal growth of various cell types. Angiotensin II (Ang II) is one of the most potent inducers of oxidative stress in the vasculature. The molecular events involved in Ang II-induced proliferation of vascular smooth muscle cells (VSMCs) are ... More
Human cytomegalovirus-encoded interleukin-10 homolog inhibits maturation of dendritic cells and alters their functionality.
AuthorsChang WL, Baumgarth N, Yu D, Barry PA,
JournalJ Virol
PubMed ID15280480
Interleukin-10 (IL-10) suppresses the maturation and cytokine production of dendritic cells (DCs), key regulators of adaptive immunity, and prevents the activation and polarization of naïve T cells towards protective gamma interferon-producing effectors. We hypothesized that human cytomegalovirus (HCMV) utilizes its viral IL-10 homolog (cmvIL-10) to attenuate DC functionality, thereby subverting ... More
Protocol to generate human liver spheroids to study liver fibrosis induced by metabolic stress.
Currently, there is no effective treatment for obesity and alcohol-associated liver diseases, partially due to the lack of translational human models. Here, we present a protocol to generate 3D human liver spheroids that contain all the liver cell types and mimic âlivers in a dish.â We describe strategies to induce ... More
Shedding of the Interleukin-6 (IL-6) Receptor (gp80) Determines the Ability of IL-6 to Induce gp130 Phosphorylation in Human Osteoblasts.
'Human osteoblasts produce interleukin-6 (IL-6) and respond to IL-6 in the presence of soluble IL-6 receptor (sIL-6R), but the cell surface expression of IL-6R and the mechanism of sIL-6R production are largely unknown. Three different human osteoblast-like cell lines (MG-63, HOS, and SaOS-2) and bone marrow-derived primary human osteoblasts expressed ... More
Active nuclear import and export pathways regulate E2F-5 subcellular localization.
'Epidermal keratinocyte differentiation is accompanied by differential regulation of E2F genes, including up-regulation of E2F-5 and its concomitant association with the retinoblastoma family protein p130. This complex appears to play a role in irreversible withdrawal from the cell cycle in differentiating keratinocytes. We now report that keratinocyte differentiation is also ... More
Molecular cloning and functional analysis of the promoter of the human squalene synthase gene.
Authors Guan G; Jiang G; Koch R L; Shechter I;
JournalJ Biol Chem
PubMed ID7665618
'We have cloned and characterized the 5''-flanking region of the gene encoding human squalene synthase. We report here the promoter activity of successively 5''-truncated sections of a 1 kilobase of this region by fusing it to the coding region of a luciferase reporter gene. DNA segments of 200 base pairs ... More
Efficacy of acetaminophen in skin B16-F0 melanoma tumor-bearing C57BL/6 mice.
AuthorsVad NM, Kudugunti SK, Graber D, Bailey N, Srivenugopal K, Moridani MY
JournalInt J Oncol
PubMed ID19513568
'Previously, we reported that acetaminophen (APAP) showed selective toxicity towards melanoma cell lines. In the current study, we investigated further the role of tyrosinase in APAP toxicity in SK-MEL-28 melanoma cells in the presence of a short hairpin RNA (shRNA) plasmid, silencing tyrosinase gene. Results from tyrosinase shRNA experiments showed ... More
c-Src-dependent transcriptional activation of TFII-I.
Authors Cheriyath Venugopalan; Desgranges Zana Patrick; Roy Ananda L;
JournalJ Biol Chem
PubMed ID11934902
'TFII-I is a multifunctional transcription factor that is also involved in signal transduction. Here we show that TFII-I undergoes a c-Src-dependent tyrosine phosphorylation on tyrosine residues 248 and 611 and translocates to the nucleus in response to growth factor signaling. Tyrosine-phosphorylated nuclear TFII-I activates a stably integrated c-fos reporter gene. ... More
Defective ureagenesis in mice carrying a liver-specific disruption of hepatocyte nuclear factor 4alpha (HNF4alpha ). HNF4alpha regulates ornithine transcarbamylase in vivo.
Authors Inoue Yusuke; Hayhurst Graham P; Inoue Junko; Mori Masataka; Gonzalez Frank J;
JournalJ Biol Chem
PubMed ID11994307
'Hepatocyte nuclear factor 4alpha (HNF4alpha) regulates the expression of many genes preferentially expressed in liver. HNF4alpha-null mice die during embryogenesis precluding the analysis of its function in the adult. To circumvent this problem, liver-specific HNF4alpha-null mice were produced. Mice lacking hepatic HNF4alpha expression exhibited increased serum ammonia and reduced serum ... More
Regulation of Jak kinases by intracellular leptin receptor sequences.
AuthorsKloek C, Haq AK, Dunn SL, Lavery HJ, Banks AS, Myers MG,
JournalJ Biol Chem
PubMed ID12196522
'Leptin signals the status of body energy stores via the leptin receptor (LR), a member of the Type I cytokine receptor family. Type I cytokine receptors mediate intracellular signaling via the activation of associated Jak family tyrosine kinases. Although their COOH-terminal sequences vary, alternatively spliced LR isoforms (LRa-LRd) share common ... More
The circadian regulatory proteins BMAL1 and cryptochromes are substrates of casein kinase Iepsilon.
Authors Eide Erik J; Vielhaber Erica L; Hinz William A; Virshup David M;
JournalJ Biol Chem
PubMed ID11875063
'The serine/threonine protein kinase casein kinase I epsilon (CKIepsilon) is a key regulator of metazoan circadian rhythm. Genetic and biochemical data suggest that CKIepsilon binds to and phosphorylates the PERIOD proteins. However, the PERIOD proteins interact with a variety of circadian regulators, suggesting the possibility that CKIepsilon may interact with ... More
Foam cell formation inhibits growth of Chlamydia pneumoniae but does not attenuate Chlamydia pneumoniae-induced secretion of proinflammatory cytokines.
Authors Blessing Erwin; Kuo Cho-Chou; Lin Tsun-Mei; Campbell Lee Ann; Bea Florian; Chesebro Brian; Rosenfeld Michael E;
JournalCirculation
PubMed ID11997286
'BACKGROUND: It has not yet been determined whether lipid-loaded macrophages (foam cells), a major cellular component of atherosclerotic lesions, have the capacity to support growth of Chlamydia pneumoniae and be activated to secrete proinflammatory cytokines in response to C pneumoniae infection. METHODS AND RESULTS: Lipid loading of RAW 264.7 cells ... More
Plaque production by the polyoma virus.
AuthorsDULBECCO R, FREEMAN G,
JournalVirology
PubMed ID13669362
Describes the original formulation of DMEM (Dulbecco’s Modified Eagle Medium), used for plaque production by the polyoma virus.
Signal-dependent regulation of splicing via phosphorylation of Sam68.
AuthorsMatter N, Herrlich P, König H,
JournalNature
PubMed ID12478298
Evolution of human organismal complexity from a relatively small number of genes--only approximately twice that of worm or fly--is explained mainly by mechanisms generating multiple proteins from a single gene, the most prevalent of which is alternative pre-messenger-RNA splicing. Appropriate spatial and temporal generation of splice variants demands that alternative ... More
The p85 regulatory subunit controls sequential activation of phosphoinositide 3-kinase by Tyr kinases and Ras.
AuthorsJimenez C, Hernandez C, Pimentel B, Carrera AC,
JournalJ Biol Chem
PubMed ID12196526
Class IA phosphoinositide 3-kinase (PI3K) is a heterodimer composed of a p85 regulatory and a p110 catalytic subunit that regulates a variety of cell responses, including cell division and survival. PI3K is activated following Tyr kinase stimulation and by Ras. We found that the C-terminal region of p85, including the ... More
Retrovirus-derived vectors are overwhelmingly preferred over other methods for ex vivo gene therapy because they provide permanent integration of foreign genes into cellular DNA. In comparison, cationic lipids mediate efficent gene transfer, but expression is transient. When we combined cationic lipids with retrovirus particles we obtained a significant enhancement of ... More
Extracellular mRNA induces dendritic cell activation by stimulating tumor necrosis factor-alpha secretion and signaling through a nucleotide receptor.
We previously demonstrated that dendritic cell (DC) pulsing with antigen-encoded mRNA resulted in the loading of both major histocompatibility complex class I and II antigen presentation pathways and the delivery of an activation signal. Coculture of mRNA-pulsed DC with T cells led to the induction of a potent primary immune ... More
The calcium sensor protein visinin-like protein-1 modulates the surface expression and agonist sensitivity of the alpha 4beta 2 nicotinic acetylcholine receptor.
Authors Lin Lin; Jeanclos Elisabeth M; Treuil Magdalen; Braunewell Karl-Heinz; Gundelfinger Eckart D; Anand Rene;
JournalJ Biol Chem
PubMed ID12202488
The calcium sensor protein visinin-like protein-1 (VILIP-1) was isolated from a brain cDNA yeast two-hybrid library using the large cytoplasmic domain of the alpha4 subunit as a bait. VILIP-1 is a myristoylated calcium sensor protein that contains three functional calcium binding EF-hand motifs. The alpha4 subunit residues 302-339 were found ... More
Dystrophin deficiency markedly increases enterovirus-induced cardiomyopathy: a genetic predisposition to viral heart disease.
Authors Xiong Dingding; Lee Gil-Hwan; Badorff Cornel; Dorner Andrea; Lee Sang; Wolf Paul; Knowlton Kirk U;
JournalNat Med
PubMed ID12118246
Both enteroviral infection of the heart and mutations in the dystrophin gene can cause cardiomyopathy. Little is known, however, about the interaction between genetic and acquired forms of cardiomyopathy. We previously demonstrated that the enteroviral protease 2A cleaves dystrophin; therefore, we hypothesized that dystrophin deficiency would predispose to enterovirus-induced cardiomyopathy. ... More
L-phenylalanine and NPS R-467 synergistically potentiate the function of the extracellular calcium-sensing receptor through distinct sites.
Authors Zhang Zaixiang; Jiang Yongfeng; Quinn Stephen J; Krapcho Karen; Nemeth Edward F; Bai Mei;
JournalJ Biol Chem
PubMed ID12114500
The extracellular calcium (Ca(2+)(o))-sensing receptor (CaR) can be potentiated by allosteric activators including calcimimetics and l-amino acids. In this study, we found that many mutations had differential effects on the functional modulation of the CaR by these two allosteric activators, supporting the idea that these modulators act through distinct sites. ... More
Identification of a mitochondrial target of thiazolidinedione insulin sensitizers (mTOT)--relationship to newly identified mitochondrial pyruvate carrier proteins.
AuthorsColca JR, McDonald WG, Cavey GS, Cole SL, Holewa DD, Brightwell-Conrad AS, Wolfe CL, Wheeler JS, Coulter KR, Kilkuskie PM, Gracheva E, Korshunova Y, Trusgnich M, Karr R, Wiley SE, Divakaruni AS, Murphy AN, Vigueira PA, Finck BN, Kletzien RF
Journal
PubMed ID23690925
Thiazolidinedione (TZD) insulin sensitizers have the potential to effectively treat a number of human diseases, however the currently available agents have dose-limiting side effects that are mediated via activation of the transcription factor PPAR?. We have recently shown PPAR?-independent actions of TZD insulin sensitizers, but the molecular target of these ... More
Influence of second and third cytoplasmic loops on binding, internalization, and coupling of chimeric bombesin/m3 muscarinic receptors.
Authors Tseng M J; Coon S; Stuenkel E; Struk V; Logsdon C D;
JournalJ Biol Chem
PubMed ID7629092
In order to investigate the molecular basis for differences in the characteristics of bombesin (Bn) and m3 muscarinic cholinergic (m3 ACh) receptors, chimeric Bn receptors possessing cytoplasmic domains from the m3 ACh receptor were produced. The receptors were expressed in CHO-K1 cells and binding, structural, and signal transduction characteristics were ... More
Identification of domains in human beta-hexosaminidase that determine substrate specificity.
Authors Pennybacker M; Liessem B; Moczall H; Tifft C J; Sandhoff K; Proia R L;
JournalJ Biol Chem
PubMed ID8663217
The lysosomal beta-hexosaminidases are dimers composed of alpha and beta subunits. beta-Hexosaminidase A (alphabeta) is a heterodimer, whereas hexosaminidase B (betabeta) and S (alphaalpha) are homodimers. Although containing a high degree of amino acid identity, each subunit expresses a unique active site that can be distinguished by a differential ability ... More
Targeted disruption of spermidine/spermine N1-acetyltransferase gene in mouse embryonic stem cells. Effects on polyamine homeostasis and sensitivity to polyamine analogues.
We have generated mouse embryonic stem cells with targeted disruption of spermidine/spermine N(1)-acetyltransferase (SSAT) gene. The targeted cells did not contain any inducible SSAT activity, and the SSAT protein was not present. The SSAT-deficient cells proliferated normally and appeared to maintain otherwise similar polyamine pools as did the wild-type cells, ... More
Mutational analysis of the insulin-like growth factor I prohormone processing site.
Authors Duguay S J; Lai-Zhang J; Steiner D F;
JournalJ Biol Chem
PubMed ID7615562
Insulin-like growth factor I (IGF-I) is a mitogenic peptide that is produced in most tissues and cell lines and plays an important role in embryonic development and postnatal growth. IGF-I is initially synthesized as a prohormone precursor that is converted to mature IGF-I by endoproteolytic removal of the carboxyl-terminal E-domain. ... More
Brief exposure to high-dose transforming growth factor-beta1 enhances periosteal chondrogenesis in vitro: a preliminary report.
BACKGROUND: Articular cartilage has limited potential for repair. There have been various attempts aimed at improving the repair process in articular cartilage. Transforming growth factor-beta1 (TGF-beta1) has a stimulatory effect on chondrogenesis in periosteal explants. The purpose of the present study was to determine the effect of brief exposures (i.e., ... More
A 2-D liquid separations/mass mapping method for interlysate comparison of ovarian cancers.
Authors Kachman Maureen T; Wang Haixing; Schwartz Donald R; Cho Kathleen R; Lubman David M;
JournalAnal Chem
PubMed ID11985308
A two-dimensional liquid phase separation of proteins from whole cell lysates coupled on-line to an electrospray-ionization time-of-flight (ESI-TOF) mass spectrometer (MS) is used to map the protein content of ovarian surface epithelial cells (OSE) and an ovarian carcinoma-derived cell line (ES2). The two dimensions involve the use of liquid isoelectric ... More