Adhesion G protein-coupled receptors (aGPCRs) constitute the second largest GPCR subfamily. GPR56/ADGRG1 is a member of the ADGRG subgroup of aGPCRs. Although GPR56 is best known for its pivotal role in the cerebral cortical development, it is also important for tumor progression. Numerous studies have revealed that GPR56 is expressed in various cancer types with a role in cancer cell adhesion, migration and metastasis. In a recent study, we found that the immobilized GPR56-specific CG4 antibody enhanced IL-6 production and migration ability of melanoma cells. In this review, we will summarize the current understanding of GPR56 function and discuss the activation and signaling mechanisms of GPR56 in melanoma cells.

Neutrophils play essential anti-microbial and inflammatory roles in host defense, however, their activities require tight regulation as dysfunction often leads to detrimental inflammatory and autoimmune diseases. Here we show that the adhesion molecule GPR97 allosterically activates CD177-associated membrane proteinase 3 (mPR3), and in conjugation with several protein interaction partners leads to neutrophil activation in humans. Crystallographic and deletion analysis of the GPR97 extracellular region identified two independent mPR3-binding domains. Mechanistically, the efficient binding and activation of mPR3 by GPR97 requires the macromolecular CD177/GPR97/PAR2/CD16b complex and induces the activation of PAR2, a G protein-coupled receptor known for its function in inflammation. Triggering PAR2 by the upstream complex leads to strong inflammatory activation, prompting anti-microbial activities and endothelial dysfunction. The role of the complex in pathologic inflammation is underscored by the finding that both GPR97 and mPR3 are upregulated on the surface of disease-associated neutrophils. In summary, we identify a PAR2 activation mechanism that directs neutrophil activation, and thus inflammation. The PR3/CD177/GPR97/PAR2/CD16b protein complex, therefore, represents a potential therapeutic target for neutrophil-mediated inflammatory diseases.

Salmonella enterica serovars Choleraesuis and Typhimurium are among the non-typhoid Salmonella serovars that are important zoonotic pathogens. In clinical observation, S. Typhimurium typically causes diarrheal diseases; however, S. Choleraesuis shows high predilection to cause bacteremia. The mechanism why S. Choleraesuis is more invasive to humans remains unknown. In this study, we compared the S. Typhimurium LT2 and S. Choleraesuis SC-B67 proteomes through stable isotope labeling of amino acid in cell culture (SILAC). In SILAC, the expression of many virulence proteins in two type III secretion systems (T3SSs) were significantly higher in S. Choleraesuis than in S. Typhimurium. Similar differences were also found at the transcriptional level. Compared to S. Typhimurium, S. Choleraesuis showed a higher penetration level to Caco-2 (>100-fold) and MDCK (>10-fold) monolayers. In mice after oral challenge, the invasion of spleen and liver was also higher in S. Choleraesuis than in S. Typhimurium. The transcription of hilD in S. Choleraesuis was increased in physiological (1 mM) or high (10 mM) concentrations of Mg2+, but not in low (8 μM) concentration. We conclude that S. Choleraesuis showed hyperinvasiveness in cellular as well as mouse models due to hyperexpression of T3SS genes.

The genus Salmonella contains more than 2500 serovars. While most cause the self-limiting gastroenteritis, a few serovars can elicit typhoid fever, a severe systemic infection. S. enterica subsp. enterica serovar Typhimurium and S. Typhi are the representatives of the gastroenteritis and typhoid fever types of Salmonella. In this study, we adopted Stable Isotope Labeling with Amino acids in Cell culture (SILAC) technology to quantitatively compare the proteomes of the two serovars. We found several proteins with serovar-specific expression, which could be developed as new biomarkers for clinical serotype diagnosis. We found that flagella and chemotaxis genes were down-regulated in S. Typhi in comparison with S. Typhimurium. We attributed this observation to the fact that the smooth cellular structure of S. Typhi may better fit its systemic lifestyle. Instead of known virulence factors that were located within Salmonella Pathogenecity Islands, a number of core genes, which were involved in metabolism and transport of carbohydrates and amino acids, showed differential expression between the two serovars. Further studies on the roles of these differentially-expressed genes in the pathogenesis should be undertaken.

FAM46A belongs to the FAM46 subfamily of the nucleotidyltransferase-fold superfamily and is predicted to be a non-canonical poly(A) polymerase. FAM46A has been linked to several human disorders including retinitis pigmentosa, bone abnormality, cancer, and obesity. However, its molecular and functional characteristics are largely unknown. We herein report that FAM46A is expressed in cells of the hematopoietic system and plays a role in hemin-induced hemoglobinization. FAM46A is a nucleocytoplasmic shuttle protein modified by Tyr-phosphorylation only in the cytosol, where it is closely associated with ER. On the other hand, it is located proximal to the chromatin regions of active transcription in the nucleus. FAM46A is a cell cycle-dependent poly-ubiquitinated short-lived protein degraded mostly by proteasome and its overexpression inhibits cell growth and promotes hemin-induced hemoglobinization in K562 cell. Site-directed mutagenesis experiments confirm the non-canonical poly(A) polymerase activity of FAM46A is essential for enhanced hemin-induced hemoglobinization. In summary, FAM46A is a novel poly(A) polymerase that functions as a critical intracellular modulator of hemoglobinization.

GPR56/ADGRG1, a member of the adhesion G protein‐coupled receptor family, is important for tumor progression. The role of GPR56 in melanoma development is unresolved. Here, we showed that GPR56 receptor ligation by immobilized CG4 mAb in melanoma cell enhanced the production of IL‐6, which promoted melanoma cell migration and invasion. siRNA‐mediated Gα12/13 knockdown significantly reduced CG4‐induced IL‐6 production in melanoma cell. Likewise, the IL‐6 level was decreased in melanoma cells expressing the dominant negative RhoA N19, treated with exoenzyme C3, or the ROCK inhibitors (Y27632 and H1152). We conclude that GPR56 activation‐induced IL‐6 production is mediated through the Gα12/13‐RhoA‐ROCK signaling pathway. IL‐6 in turn promotes melanoma cell migration and invasion. Our results indicate a critical role of GPR56 in melanoma progression.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Neutrophils play essential anti-microbial and inflammatory roles in host defense, however their activities are tightly regulated as neutrophil dysfunction often leads to detrimental inflammatory and autoimmune diseases. Here, we identified a novel PR3/CD177/GPR97/PAR2/CD16b interactome as a critical modulator of neutrophil activation. Using multiple approaches, we deorphanized GPR97, a human neutrophil-restricted adhesion G protein-coupled receptor (aGPCR), as the interacting protein and allosteric activator of CD177-associated membrane proteinase 3 (mPR3). Structural and deletion analysis of the GPR97 extracellular region disclosed two independent mPR3-binding domains. The efficient binding and activation of mPR3 by GPR97 required a macromolecular CD177/GPR97/PAR2/CD16b interactome and resulted in the transactivation of PAR2, another GPCR. GPR97-mediated PAR2 transactivation in neutrophils elicited strong inflammatory activation, triggering anti-microbial activities and endothelial dysfunction. Altogether, we identify a novel aGPCR-GPCR transactivation mechanism that directs neutrophil activation and inflammation. The PR3/CD177/GPR97/PAR2/CD16b interactome represents a potential therapeutic target for neutrophil-mediated inflammatory diseases.