JMJD5, a Jumonji C domain-containing dioxygenase, is important for embryonic development and cancer growth. Here, we show that JMJD5 is up-regulated by hypoxia and is crucial for hypoxia-induced cell proliferation. JMJD5 interacts directly with pyruvate kinase muscle isozyme (PKM)2 to modulate metabolic flux in cancer cells. The JMJD5-PKM2 interaction resides at the intersubunit interface region of PKM2, which hinders PKM2 tetramerization and blocks pyruvate kinase activity. This interaction also influences translocation of PKM2 into the nucleus and promotes hypoxia-inducible factor (HIF)-1α–mediated transactivation. JMJD5 knockdown inhibits the transcription of the PKM2–HIF-1α target genes involved in glucose metabolism, resulting in a reduction of glucose uptake and lactate secretion in cancer cells. JMJD5, along with PKM2 and HIF-1α, is recruited to the hypoxia response element site in the lactate dehydrogenase A and PKM2 loci and mediates the recruitment of the latter two proteins. Our data uncover a mechanism whereby PKM2 can be regulated by factor-binding–induced homo/heterooligomeric restructuring, paving the way to cell metabolic reprogram. Significance Cancer cells favor high rates of aerobic glycolysis, or the Warburg effect, which is mediated by a key molecule, pyruvate kinase muscle isozyme (PKM)2. PKM2 functions both as a cytosolic enzyme and a nuclear factor in tumor cells. This report shows that PKM2’s nuclear translocation is regulated by Jumonji C domain-containing dioxygenase (JMJD)5 via direct physical binding. JMJD5 hinders the PKM2 tetrameric assembly and facilitates PKM2’s nuclear translocation. Together, they modulate hypoxia-inducible factor 1α-mediated transcriptional reprogramming of metabolic genes. These results reveal a mechanism whereby PKM2’s activity can be modulated by a dioxygenase/demethylase.

Infection withHelicobacter pylori cagA-positive strains is associated with gastritis, ulcerations, and gastric cancer. CagA is translocated into infected epithelial cells by a type IV secretion system and can be tyrosine phosphorylated, inducing signal transduction and motogenic responses in epithelial cells. Cellular cholesterol, a vital component of the membrane, contributes to membrane dynamics and functions and is important in VacA intoxication and phagocyte evasion duringH. pyloriinfection. In this investigation, we showed that cholesterol extraction by methyl-β-cyclodextrin reduced the level of CagA translocation and phosphorylation. Confocal microscope visualization revealed that a significant portion of translocated CagA was colocalized with the raft marker GM1 and c-Src during infection. Moreover, GM1 was rapidly recruited into sites of bacterial attachment by live-cell imaging analysis. CagA and VacA were cofractionated with detergent-resistant membranes (DRMs), suggesting that the distribution of CagA and VacA is associated with rafts in infected cells. Upon cholesterol depletion, the distribution shifted to non-DRMs. Accordingly, the CagA-induced hummingbird phenotype and interleukin-8 induction were blocked by cholesterol depletion. Raft-disrupting agents did not influence bacterial adherence but did significantly reduce internalization activity in AGS cells. Together, these results suggest that delivery of CagA into epithelial cells by the bacterial type IV secretion system is mediated in a cholesterol-dependent manner.

The KDM4/JMJD2 Jumonji C-containing histone lysine demethylases (KDM4A–KDM4D), which selectively remove the methyl group(s) from tri/dimethylated lysine 9/36 of H3, modulate transcriptional activation and genome stability. The overexpression of KDM4A/KDM4B in prostate cancer and their association with androgen receptor suggest that KDM4A/KDM4B are potential progression factors for prostate cancer. Here, we report the crystal structure of the KDM4B·pyridine 2,4-dicarboxylic acid·H3K9me3 ternary complex, revealing the core active-site region and a selective K9/K36 site. A selective KDM4A/KDM4B inhibitor, 4, that occupies three subsites in the binding pocket is identified by virtual screening. Pharmacological and genetic inhibition of KDM4A/KDM4B significantly blocks the viability of cultured prostate cancer cells, which is accompanied by increased H3K9me3 staining and transcriptional silencing of growth-related genes. Significantly, a substantial portion of differentially expressed genes are AR-responsive, consistent with the roles of KDM4s as critical AR activators. Our results point to KDM4 as a useful therapeutic target and identify a new inhibitor scaffold.

Objectives: Helicobacter pylori resistance to antibiotics is the main factor for therapy failure, while other features remain largely unknown. The aims of this study are to investigate the relationship of antibiotic resistance and in vitro internalization activity between cure and failure isolates and to determine whether failures are associated with persistence of the same predominant strain. Methods: Fifty-three isolates from forty-seven patients (cure group, n = 31; failure group, n = 16) receiving one of two lansoprazole-based therapies before and/or after therapy were investigated. Antibiotic susceptibility was determined by Etest. Genotyping was determined by cagA, babA, vacA and RAPD analyses. Target cells of internalization assay were AGS cells. Results: Five of six paired pre- and post-treatment isolates had the same predominant genetic profiles and exhibited similarly high internalization activities. The A2143G point mutation of the 23S rRNA gene conferred clarithromycin resistance. Moreover, increased antibiotic resistance after therapy was found for these five cases. Pre-treatment isolates from the failure group (n = 11) had higher level of internalization activity than those from the cure group (n = 31) (P = 0.00005). Antibiotic-resistant strains were significantly associated with higher internalization activity than were susceptible strains (metronidazole, P < 0.005; clarithromycin, P < 0.005). Conclusions: Our results suggest that resistant H. pylori strains are associated with antibiotic resistance and superior internalization activity, protecting them against antibiotic treatment.

Summary: Helicobacter pylori infection is an aetiological cause of gastric disorders worldwide. H. pylori has been shown to assimilate and convert host cholesterol into cholesteryl glucosides (CGs) by cholesterol‐α‐glucosyltransferase encoded by capJ. Here, we show that CapJ‐deficient (ΔcapJ) H. pylori resulted in greatly reduced type IV secretion system (TFSS)‐associated activities, including the hummingbird phenotype of AGS cells, IL‐8 production, CagA translocation/phosphorylation and CagA‐mediated signalling events. Complementation of the ΔcapJ mutation with wild type cagJ or by adding CGs‐containing lysates or exogenous fluorophore‐tagged CGs reversed the mutant phenotypes. We also show that the wild‐type but not ΔcapJ H. pylori recruited raft‐associated components to sites of bacterial attachment. Fluorescence recovery after photobleaching (FRAP) analysis of AGS cells treated with fluorescence‐tagged cholesterol/CGs revealed that there was a higher proportion of CGs associated with immobile fractions. CGs‐associated membranes were also more resistant to a cold detergent extraction. Thus, we propose that CGs synthesized by H. pylori around host‐pathogen contact sites partition in detergent‐resistant membranes (DRMs), alters lateral‐phase segregation in membrane and reorganizes membrane architecture. These processes together promote the formation of a functional TFSS and H. pylori infection.

Rationale: The progression of prostate cancer (PCa) to castration-resistant PCa (CRPC) despite continuous androgen deprivation therapy is a major clinical challenge. Over 90% of patients with CRPC exhibit sustained androgen receptor (AR) signaling. KDM4B that removes the repressive mark H3K9me3/2 is a transcriptional activator of AR and has been implicated in the development of CRPC. However, the mechanisms of KDM4B involvement in CRPC remain largely unknown. Here, we sought to demonstrate the molecular pathway mediated by KDM4B in CRPC and to provide proof-of-concept evidence that KDM4B is a potential CRPC target.