G-protein-coupled receptor kinases (GRKs) are an important family of Ser/Thr kinases that specifically phosphorylate and desensitize the activated receptor in response to environmental stimulation. Here we identify p53, a key tumor suppressor, as a novel GRK substrate in vivo, revealing a previously unknown function of GRKs in regulation of genome stability. Knockdown GRK5 in osteosarcoma cells inhibits DNA damage-induced apoptosis via a p53-mediated mechanism. Furthermore, GRK5, but not GRK2 or GRK6, phosphorylates p53 at Thr-55, which promotes the degradation of p53, leading to inhibition of p53-dependent apoptotic response to genotoxic damage. Consistently, the increase of p53 and irradiation-induced apoptosis were observed in GRK5-deficient mice. These results demonstrate GRK5 as a novel kinase of p53, as well as a negative regulator of p53-mediated signal transduction. G-protein-coupled receptor (GPCR)2 kinases (GRKs) are known as a family of serine/threonine kinases that catalyze phosphorylation of seven-transmembrane-spanning G-protein-coupled receptors. They play important roles in the initiation of homologous desensitization of stimulated receptors and thus act as crucial negative regulators of GPCRs (1). GRKs have been demonstrated to phosphorylate and sequester a variety of GPCRs, including adrenergic receptor, muscarinic receptor, dopamine receptors, opioid receptors, chemokine receptor (2-6), etc.Regulation of receptors other than GPCR family by GRKs has also been shown recently by others and our laboratory (7-9). GRK2 can phosphorylate tyrosine kinase-coupled receptors, including epidermal growth factor receptor and platelet-derived growth factor receptor (7,8), and regulate 12-transmembrane receptor Patched 1 (9). Transgenic or knock-out studies have demonstrated that deletion or overexpression of GRK in mice causes dysfunction of cardiovascular, nervous, respiratory, optical, and immune systems or even embryonic lethality (3, 10 -13). In addition to their well established roles in the regulation of receptor-mediated signaling, emerging evidence indicates that GRKs are also capable of phosphorylating nonreceptor substrates, such as synuclein (14) and ␤-arrestin 1 (15). However, the physiological significance of these phosphorylation events mediated by GRKs remains unclear.Seven subtypes of GRKs (GRK1-7) have been identified to date. Members of the GRK family share highly homologous structure and similar function in the regulation of GPCR signaling (10). Most subtypes of GRKs, including GRK2, GRK3, GRK5, and GRK6, are ubiquitously expressed in most tissues (1, 16). Deletion of most individual GRK subtypes, such as GRK1, GRK3, GRK4, GRK5, GRK6, and GRK7, does not result in serious phenotype in mice (10), which indicates a redundancy of GRK subtypes in the regulation of GPCR signaling. In this regard, the functional specificity of an individual GRK subtype remains to be explored. Johnson et al. (17) and Yi et al. (18) showed recently that GRK5 contains a putative nuclear localization sequence and ...

Constitutive activation of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway occurs frequently in cancer cells and contributes to oncogenesis. Among the members of STAT family, STAT3 plays a pivotal role in the development and progression of human tumors. The STAT3-mediated signaling pathway has been recognized as a promising anticancer target. Here, we show that 17-Hydroxy-jolkinolide B (HJB), a diterpenoid from the Chinese medicinal herb Euphorbia fischeriana Steud, strongly inhibits interleukin (IL)-6-induced as well as constitutive STAT3 activation. Furthermore, we show that HJB directly targets the JAK family kinases, JAK1, JAK2, and TYK2, by inducing dimerization of the JAKs via cross-linking. Addition of DTT or glutathione prevents the JAK crosslinking and blocks the inhibitory effects of HJB on IL-6-induced STAT3 activation, suggesting that HJB may react with cystein residues of JAKs to form covalent bonds that inactivate JAKs. Liquid chromatography/mass spectrometry analysis confirmed that each HJB reacted with two thiols. The effect of HJB on the JAK/STAT3 pathway is specific as HJB has no effect on platelet-derived growth factor, epidermal growth factor, or insulin-like growth factor I signaling pathways. Finally, we show that HJB inhibits growth and induces apoptosis of tumor cells, particularly those tumor cells with constitutively activated STAT3. We propose that the natural compound HJB is a promising anticancer drug candidate as a potent STAT3 signaling inhibitor. [Cancer Res 2009;69(18):7302-10]

Antimicrobial growth promoters (AGPs) are commonly used in the livestock industry at subtherapeutic levels to improve production efficiency, which is achieved mainly through modulation of the intestinal microbiota. However, how different classes of AGPs, particularly ionophores, regulate the gut microbiota remains unclear. In this study, male Cobb broiler chickens were supplemented for 14 days with or without one of five commonly used AGPs including three classical antibiotics (bacitracin methylene disalicylate, tylosin, and virginiamycin) and two ionophores (monensin and salinomycin) that differ in antimicrobial spectrum and mechanisms. Deep sequencing of the V3-V4 region of the bacterial 16S rRNA gene revealed that two ionophores drastically reduced a number of rare bacteria resulting in a significant decrease in richness and a concomitant increase in evenness of the cecal microbiota, whereas three antibiotics had no obvious impact. Although each AGP modulated the gut microbiota differently, the closer the antibacterial spectrum of AGPs, the more similarly the microbiota was regulated. Importantly, all AGPs had a strong tendency to enrich butyrate- and lactic acid-producing bacteria, while reducing bile salt hydrolase-producing bacteria, suggestive of enhanced metabolism and utilization of dietary carbohydrates and lipids and improved energy harvest, which may collectively be responsible for the growth-promoting effect of AGPs.

A study was conducted to investigate the effect of dietary yeast polysaccharides on some hematologic parameters and intestinal morphology of channel catfish. Channel catfish were fed diets containing yeast polysaccharides at 0 (control), 0.1, 0.2, or 0.3 % for 7 weeks. Each diet was provided to 10 channel catfish specimens (5.82 ± 0.13 g initial weight) replicated 3 times in individual 250 L fiberglass tanks. Some hematologic parameters, leukocyte phagocytic activity, and intestinal morphology were monitored. After 7 weeks of trial, 0.2 % yeast polysaccharides resulted in significantly higher (P < 0.05) monocyte numbers. Furthermore, fish fed 0.2 % yeast polysaccharide diet had higher (P < 0.05) phagocytic rate of leukocyte. And 0.3 % yeast polysaccharide enhanced (P < 0.05) phagocytic index of leukocyte. Histological evaluation showed yeast polysaccharide supplementation increased the height of intestine fold (0.1, 0.2 and 0.3 %) and the thick of muscular layers (0.2 %) in intestine (P < 0.05). In addition, 0.1 and 0.3 % yeast polysaccharide supplementation improved the number of goblet cells (P < 0.05). The results of this trial indicate that yeast polysaccharides supplementation could affect blood monocytes, improve leukocytes phagocytic activity, and the development of intestine in channel catfish.

1. A study was conducted to investigate the effects of an esterified glucomannan (EGM), a hydrated sodium calcium aluminosilicate (HSCAS) and a compound mycotoxin adsorbent (CMA) on performance, nutrient retention and meat quality in broilers fed on mould-contaminated feed. Mould-contaminated diets were prepared by replacing half of the non-contaminated maize in the basal diets with mould-contaminated maize, which contained 450·6 µg/kg of aflatoxin B1, 68·4 µg/kg of ochratoxin A and 320·5 µg/kg of T-2 toxin. 2. The mould-contaminated diet significantly decreased body weight gain (BWG) between 10 and 21 d, feed intake (FI) between 35 and 42 d, the apparent retention of crude lipid and phosphorus, and the lightness (L*) value of breast and thigh muscle. It also significantly increased the redness (a*) and yellowness (b*) value in breast muscle and the b* value in thigh muscle. 3. The addition of 0·2% HSCAS significantly increased FI between 35 and 42 d and the apparent retention of phosphorus. Supplementation with 0·1% CMA in the contaminated diet significantly improved BWG from 10 to 21 d, and increased FI from 35 to 42 d and from 10 to 42 d. CMA also significantly increased the apparent retention of crude lipid, crude protein, ash and phosphorus. All three mycotoxin-adsorbent treatments significantly improved the L* values of breast and thigh muscle when compared with the mould-contaminated group. Supplementation with 0·1% CMA in the contaminated diet significantly decreased b* value and improved tenderness in thigh muscle. 0·05% EGM significantly decreased b* value of thigh muscle compared to mould-contaminated group. 4. The results indicated that mycotoxins in contaminated feed retard growth, nutrient retention and meat quality, whereas the addition of 0·05% EGM, 0·2% HSCAS or 0·1% CMA prevents the adverse effects of mycotoxins to varying extents, with 0·1% CMA being the most effective adsorbent treatment.