The rockefeller university

Transcription-repair coupling factor (TRCF; the product of the mfd gene) is a widely conserved bacterial protein that couples DNA repair with transcription. TRCF recognizes RNA polymerase stalled at a noncoding lesion in the DNA template strand, uses the energy from ATP hydrolysis to disrupt the transcription complex, and stimulates DNA repair by recruiting UvrA, a component of the nucleotide excision repair machinery, to the site. TRCF is a large (130 kDa) multifunctional protein with a complex structure-function relationship consisting of a compact arrangement of eight structured domains linked by flexible linkers. Through a conserved, intramolecular, interdomain interaction, TRCF is held in a conformation in which its enzymatic activities (ATPase activity and DNA translocase activity) are strongly repressed. Disruption of the repressive interdomain interaction by amino acid substitutions within the interface derepresses ATPase and DNA translocase activities. In this work, we have shown that derepressed TRCF mutants are dramatically sensitized to limited proteolysis compared with repressed TRCF, pointing to an altered conformational state. Analysis of the protease cleavage sites mapped onto the structure of the repressed TRCF conformation indicates that (1) the cleavage sites tend to cluster at linkers connecting the TRCF structured domains, and (2) many of the cleavage sites sensitized in the derepressed TRCF are partially or completely buried to protease access in the repressed TRCF structure. We conclude that TRCF derepression is associated with profound conformational changes that primarily involve a reorganization of the interdomain interactions. (C) 2010 Elsevier Ltd. All rights reserved.

The identity of higher-order neurons and circuits playing an associative role to control feeding is unknown. We injected pseudorabies virus, a retrograde tracer, into masseter muscle, salivary gland, and tongue of BAC-transgenic mice expressing GFIP in specific neural populations and identified several CNS regions that project multisynaptically to the periphery. MCH and orexin neurons were identified in the lateral hypothalamus, and Nurrl and Cnr1 in the amygdala and insular/rhinal cortices. Cholera toxin beta tracing showed that insular Nurrl(+) and Cnr1(+) neurons project to the amygdala or lateral hypothalamus, respectively. Finally, we show that cortical Cnr1(+) neurons show increased Cnrl mRNA and c-Fos expression after fasting, consistent with a possible role for Cnr1(+) neurons in feeding. Overall, these studies define a general approach for identifying specific molecular markers for neurons in complex neural circuits. These markers now provide a means for functional studies of specific neuronal populations in feeding or other complex behaviors.

We studied ten individuals from eight families showing features consistent with the immuno-osseous dysplasia spondyloenchondrodysplasia. Of particular note was the diverse spectrum of autoimmune phenotypes observed in these individuals (cases), including systemic lupus erythematosus, Sjogren's syndrome, hemolytic anemia, thrombocytopenia, hypothyroidism, inflammatory myositis, Raynaud's disease and vitiligo. Haplotype data indicated the disease gene to be on chromosome 19p13, and linkage analysis yielded a combined multipoint log(10) odds (LOD) score of 3.6. Sequencing of ACP5, encoding tartrate-resistant acid phosphatase, identified biallelic mutations in each of the cases studied, and in vivo testing confirmed a loss of expressed protein. All eight cases assayed showed elevated serum interferon alpha activity, and gene expression profiling in whole blood defined a type I interferon signature. Our findings reveal a previously unrecognized link between tartrate-resistant acid phosphatase activity and interferon metabolism and highlight the importance of type I interferon in the genesis of autoimmunity.

The authors discuss one type of general forward-backward stochastic differential equations (FBSDEs) with It's stochastic delayed equations as the forward equations and anticipated backward stochastic differential equations as the backward equations. The existence and uniqueness results of the general FBSDEs are obtained. In the framework of the general FBSDEs in this paper, the explicit form of the optimal control for linearquadratic stochastic optimal control problem with delay and the Nash equilibrium point for nonzero sum differential games problem with delay are obtained.

Cdc14 phosphatase regulates multiple events during anaphase and is essential for mitotic exit in budding yeast. Cdc14 is regulated in both a spatial and temporal manner. It is sequestered in the nucleolus for most of the cell cycle by the nucleolar protein Net1 and is released into the nucleus and cytoplasm during anaphase. To identify novel binding partners of Cdc14, we used affinity purification of Cdc14 and mass spectrometric analysis of interacting proteins from strains in which Cdc14 localization or catalytic activity was altered. To alter Cdc14 localization, we used a strain deleted for NET1, which causes full release of Cdc14 from the nucleolus. To alter Cdc14 activity, we generated mutations in the active site of Cdc14 (C283S or D253A), which allow binding of substrates, but not dephosphorylation, by Cdc14. Using this strategy, we identified new interactors of Cdc14, including multiple proteins involved in mitotic events. A subset of these proteins displayed increased affinity for catalytically inactive mutants of Cdc14 compared with the wild-type version, suggesting they are likely substrates of Cdc14. We have also shown that several of the novel Cdc14-interacting proteins, including Kar9 (a protein that orients the mitotic spindle) and Bni1 and Bnr1 (formins that nucleate actin cables and may be important for actomyosin ring contraction) are specifically dephosphorylated by Cdc14 in vitro and in vivo. Our findings suggest the dephosphorylation of the formins may be important for their observed localization change during exit from mitosis and indicate that Cdc14 targets proteins involved in wide-ranging mitotic events.

We present a search for a new heavy charged vector boson W' decaying to an electron- neutrino pair in p (p) over bar collisions at a center-of-mass energy of 1.96 TeV. The data were collected with the CDF II detector and correspond to an integrated luminosity of 5: 3 fb(-1). No significant excess above the standard model expectation is observed and we set upper limits on sigma . B(W' -> e nu). Assuming standard model couplings to fermions and the neutrino from the W' boson decay to be light, we exclude a W' boson with mass less than 1.12 TeV/c(2) at the 95% confidence level.

The detection of chemosensory stimuli by the sensory neurons of the mouse vomeronasal organ (VNO) is mainly mediated by seven-transmembrane receptors that are encoded by two large gene repertoires, V1R and V2R. The mouse genome contains 122 intact V2R genes, which can be grouped in four families by sequence homology: families A, B. and D (115 genes), and family C (7 genes). Vomeronasal sensory neurons (VSNs) in the basal layer of the VNO epithelium coexpress two V2R genes in non-random combinations: one family-ABD V2R gene together with one family-C V2R gene, such as Vmn2r1 (29% of basal VSNs) or Vmn2r2 (52%). This coordinated coexpression may contribute to the highly specialized sensory response profiles of VSNs, for instance by heterodimerization of a family-ABD with a family-C V2R. The mechanisms that regulate this coordinated cooexpression of two V2R genes per basal VSN are not understood. Among possible models are a sequential and dependent model of expression; a model of random combinations of expression followed by cellular selection of VSNs with appropriate combinations; and a model of direct coordination of gene expression by another gene family such as genes encoding transcription factors. Here, we describe two novel mouse strains with targeted mutations in the family-ABD V2R gene V2rf2 that begin to provide insight into this problem. We observe that the great majority of VSNs that express intact V2rf2 coexpress Vmn2r1 immunoreactivity, and that the percentage of Vmn2r1 coexpression increases from 3 to 10 wk. Having established this tight coexpression of V2rf2 with Vmn2r1, we then asked if it is maintained when the coding sequence of V2rf2 is deleted. We find that the number of VSNs expressing a locus with a targeted deletion in the coding sequence of V2rf2 that is likely a null mutation, is similar to the number of VSNs that express intact V2rf2. But 25% of these VSNs coexpress another family-ABD V2R, which is consistent with the absence of negative feedback from the mutated V2rf2 locus. Interestingly, 9.5% of VSNs expressing the targeted deletion of V2rf2 now coexpress Vmn2r2. Finally, the marginal region of the VNO epithelium, where immature VSNs are concentrated, has more RNA of family-ABD V2R genes than of family-C genes in postnatal wild-type mice. Our results are most consistent with the sequential and dependent model for the coordinated coexpression of two V2R genes per basal VSN. (C) 2010 Elsevier Inc. All rights reserved.

Odorant receptor (OR) genes constitute with 1200 members the largest gene family in the mouse genome. A mature olfactory sensory neuron (OSN) is thought to express just one OR gene, and from one allele. The cell bodies of OSNs that express a given OR gene display a mosaic pattern within a particular region of the main olfactory epithelium. The mechanisms and cis-acting DNA elements that regulate the expression of one OR gene per OSN - OR gene choice - remain poorly understood. Here, we describe a reporter assay to identify minimal promoters for OR genes in transgenic mice, which are produced by the conventional method of pronuclear injection of DNA. The promoter transgenes are devoid of an OR coding sequence, and instead drive expression of the axonal marker tau-beta-galactosidase. For four mouse OR genes (M71, M72, MOR23, and P3) and one human OR gene (hM72), a mosaic, OSN-specific pattern of reporter expression can be obtained in transgenic mice with contiguous DNA segments of only similar to 300 bp that are centered around the transcription start site (TSS). The -150 bp region upstream of the TSS contains three conserved sequence motifs, including homeodomain (HD) binding sites. Such HD binding sites are also present in the H and P elements, DNA sequences that are known to strongly influence OR gene expression. When a 19mer encompassing a HD binding site from the P element is multimerized nine times and added upstream of a MOR23 minigene that contains the MOR23 coding region, we observe a dramatic increase in the number of transgene-expressing founders and lines and in the number of labeled OSNs. By contrast, a nine times multimerized 19mer with a mutant HD binding site does not have these effects. We hypothesize that HD binding sites in the H and P elements and in OR promoters modulate the probability of OR gene choice. (C) 2010 Elsevier Inc. All rights reserved.

Psoriasis vulgaris is a chronic skin disease in which our understanding of the pathogenesis has substantially grown in recent years. Our current appreciation of the role of the immune system is that it plays a necessary and driving role in the disease process. Investigations into the genetics of psoriasis has spurred further examinations into the contributions of immune mediators such as IL-23, IL-17, IL-22, and TNF as well as cellular mediators including a variety of dendritic cell populations of the skin and the growing number of T cell types, including the Th17 and Th22 subsets. Investigations into how these soluble and cellular elements interact with each other and the skin and form complex signal circuits to engender the psoriasis phenotype is starting to become elucidated. Furthermore, these recent advances have been fruitful in leading to the development of new classes of biologic therapeutics that are remarkably effective in halting the disease process.

Fish oils containing both EPA and DHA have been shown to have beneficial cardiovascular effects, but less is known about the independent effects of DHA. This study was designed to examine the effects of DHA on plasma lipid and lipoprotein concentrations and other biomarkers of cardiovascular risk in the absence of weight loss. In this randomized, controlled, double-blind trial, 36 overweight or obese adults were treated with 2 g/d of algal DHA or placebo for 4.5 mo. Markers of cardiovascular risk were assessed before and after treatment. In the DHA-supplemented group, the decrease in mean VLDL particle size (P <= 0.001) and increases in mean LDL (P <= 0.001) and HDL (P <= 0.001) particle sizes were significantly greater than changes in the placebo group. DHA supplementation also increased the concentrations of large LDL (P <= 0.001) and large HDL particles (P = 0.001) and decreased the concentrations of small LDL (P = 0.009) and medium HDL particles (P = 0.001). As calculated using NMR-derived data, DHA supplementation reduced VLDL TG (P = 0.009) and total TG concentrations (P = 0.006). Plasma IL-10 increased with DHA supplementation to a greater extent than placebo (P = 0.021), but no other significant changes were observed in glucose metabolism, insulin sensitivity, blood pressure, or markers of inflammation with DHA. In summary, DHA supplementation resulted in potentially beneficial changes in some., markers of cardiometabolic risk, whereas other markers were unchanged. J. Nutr. 141: 207-213, 2011.