Regulation of Human Formyl Peptide Receptor 1 Synthesis: Role of Single Nucleotide Polymorphisms, Transcription Factors, and Inflammatory Mediators

The gene encoding the human formyl peptide receptor 1 (FPR1) is heterogeneous, containing numerous single nucleotide polymorphisms (SNPs). Here, we examine the effect of these SNPs on gene transcription and protein translation. We also identify gene promoter sequences and putative FPR1 transcription factors. To test the effect of codon bias and codon pair bias on FPR1 expression, four FPR1 genetic variants were expressed in human myeloid U937 cells fused to a reporter gene encoding firefly luciferase. No significant differences in luciferase activity were detected, suggesting that the translational regulation and protein stability of FPR1 are modulated by factors other than the SNP codon bias and the variant amino acid properties. Deletion and mutagenesis analysis of the FPR1 promoter showed that a CCAAT box is not required for gene transcription. A −88/41 promoter construct resulted in the strongest transcriptional activity, whereas a −72/41 construct showed large reduction in activity. The region between −88 and −72 contains a consensus binding site for the transcription factor PU.1. Mutagenesis of this site caused significant reduction in reporter gene expression. The PU.1 binding was confirmed in vivo by chromatin immunoprecipitation, and the binding to nucleotides −84 to −76 (TTCCTATTT) was confirmed in vitro by an electrophoretic mobility shift assay. Thus, similar to many other myeloid genes, FPR1 promoter activity requires PU.1. Two single nucleotide polymorphisms at −56 and −54 did not significantly affect FPR1 gene expression, despite differences in binding of transcription factor IRF1 in vitro. Inflammatory mediators such as interferon-γ, tumor necrosis factor-α, and lipopolysaccharide did not increase FPR1 promoter activity in myeloid cells, whereas differentiation induced by DMSO and retinoic acid enhanced the activity. This implies that the expression of FPR1 in myeloid cells is developmentally regulated, and that the differentiated cells are equipped for immediate response to microbial infections

Seeing the other side: Perspective taking and the moderation of extremity

Recognizing the reasonableness of others' positions is important for conflict reduction, but is notoriously hard. We tested a perspective-taking approach to decreasing attitude entrenchment. Participants were held accountable in a task in which they wrote about a controversial issue from the perspective of a partner with an opposing viewpoint. This approach was effective at changing views on controversial issues-in Study 1 on weight discrimination, an issue participants were unlikely to have thought much about, and in Study 2 on abortion, where beliefs tend to be more deeply held. Studies 3 and 4 showed this change only took place under conditions where participants met the individual with an opposing view in person, and where that individual would see the perspective-taking effort. These results suggest that it is possible to reduce attitude entrenchment by encouraging people to think about the opposing perspective of another, as long as there is real contact and accountability

Tuning recombinant protein expression to match secretion capacity

Abstract Background The secretion of recombinant disulfide-bond containing proteins into the periplasm of Gram-negative bacterial hosts, such as E. coli, has many advantages that can facilitate product isolation, quality and activity. However, the secretion machinery of E. coli has a limited capacity and can become overloaded, leading to cytoplasmic retention of product; which can negatively impact cell viability and biomass accumulation. Fine control over recombinant gene expression offers the potential to avoid this overload by matching expression levels to the host secretion capacity. Results Here we report the application of the RiboTite gene expression control system to achieve this by finely controlling cellular expression levels. The level of control afforded by this system allows cell viability to be maintained, permitting production of high-quality, active product with enhanced volumetric titres. Conclusions The methods and systems reported expand the tools available for the production of disulfide-bond containing proteins, including antibody fragments, in bacterial hosts

A Cognitive Neuroscience Perspective On Bimanual Coordination And Interference

We argue that bimanual coordination and interference depends critically on how these actions are represented on a cognitive level. We first review the literature on spatial interactions, focusing on the difference between movements directed at visual targets and movements cued symbolically. Interactions manifest during response planning are limited to the latter condition. These results suggest that interactions in the formation of the trajectories of the two hands are associated with processes involved in response selection, rather than interactions in the motor system. Neuropsychological studies involving callosotomy patients argue that these interactions arise from transcallosal interactions between cortically-based spatial codes. The second half of the chapter examines temporal constraints observed in bimanual movements. We propose that most bimanual movements are marked by a common event structure, an explicit representation that ensures temporal coordination of the movements. The translation of an abstract event structure into a movement with a particular timing pattern is associated with cerebellar function, although the resulting temporal coupling during bimanual movements may be due to the operation of other subcortical mechanisms. For rhythmic movements that do not entail an event structure, timing may be an emergent property. Under such conditions, both spatial and temporal coupling can be absent. The emphasis on abstract levels of constraint makes clear that limitations in bimanual coordination overlap to a considerable degree with those observed in other domains of cognition