Regulation of Tumor Suppressor p53 and HCT116 Cell Physiology by Histone Demethylase JMJD2D/KDM4D

JMJD2D, also known as KDM4D, is a histone demethylase that removes methyl moieties from lysine 9 on histone 3 and from lysine 26 on histone 1.4. Here, we demonstrate that JMJD2D forms a complex with the p53 tumor suppressor in vivo and interacts with the DNA binding domain of p53 in vitro. A luciferase reporter plasmid driven by the promoter of p21, a cell cycle inhibitor and prominent target gene of p53, was synergistically activated by p53 and JMJD2D, which was dependent on JMJD2D catalytic activity. Likewise, overexpression of JMJD2D induced p21 expression in U2OS osteosarcoma cells in the absence and presence of adriamycin, an agent that induces DNA damage. Furthermore, downregulation of JMJD2D inhibited cell proliferation in wild-type and even more so in p53−/− HCT116 colon cancer cells, suggesting that JMJD2D is a pro-proliferative molecule. JMJD2D depletion also induced more strongly apoptosis in p53−/− compared to wild-type HCT116 cells. Collectively, our results demonstrate that JMJD2D can stimulate cell proliferation and survival, suggesting that its inhibition may be helpful in the fight against cancer. Furthermore, our data imply that activation of p53 may represent a mechanism by which the pro-oncogenic functions of JMJD2D become dampened

FUS/TLS Is a Co-Activator of Androgen Receptor in Prostate Cancer Cells

Androgen receptor (AR) is a member of the nuclear receptor family of transcription factors. Upon binding to androgens, AR becomes transcriptionally active to regulate the expression of target genes that harbor androgen response elements (AREs) in their promoters and/or enhancers. AR is essential for the growth and survival of prostate cancer cells and is therefore a target for current and next-generation therapeutic modalities against prostate cancer. Pathophysiologically relevant protein-protein interaction networks involving AR are, however, poorly understood. In this study, we identified the protein FUsed/Translocated in LipoSarcoma (FUS/TLS) as an AR-interacting protein by co-immunoprecipitation of endogenous proteins in LNCaP human prostate cancer cells. The hormonal response of FUS expression in LNCaP cells was shown to resemble that of other AR co-activators. FUS displayed a strong intrinsic transactivation capacity in prostate cancer cells when tethered to basal promoters using the GAL4 system. Chromatin immunoprecipitation experiments showed that FUS was recruited to ARE III of the enhancer region of the PSA gene. Data from ectopic overexpression and “knock-down” approaches demonstrated that AR transcriptional activity was enhanced by FUS. Depletion of FUS reduced androgen-dependent proliferation of LNCaP cells. Thus, FUS is a novel co-activator of AR in prostate cancer cells

Effects of a parental program for preventing underage drinking - The NGO program strong and clear

<p>Abstract</p> <p>Background</p> <p>The present study is an evaluation of a 3-year parental program aiming to prevent underage drinking. The intervention was implemented by a non-governmental organization and targeted parents with children aged 13-16 years old and included recurrent activities during the entire period of secondary school. The program consisted of four different types of group and self-administered activities: parent meetings, family dialogues, friend meetings, and family meetings.</p> <p>Methods</p> <p>A quasi-experimental design was used following parents and children with questionnaires during the three years of secondary school. The analytic sample consisted of 509 dyads of parents and children. Measures of parental attitudes and behaviour concerning underage drinking and adolescents' lifetime alcohol consumption and drunkenness were used. Three socio-demographic factors were included: parental education, school, and gender of the child. A Latent Growth Modelling (LGM) approach was used to examine changes in parental behaviour regarding youth drinking and in young people's drinking behaviour. To test for the pre-post test differences in parental attitudes repeated measures ANOVA were used.</p> <p>Results</p> <p>The results showed that parents in the program maintained their restrictive attitude toward underage drinking to a higher degree than non-participating parents. Adolescents of participants were on average one year older than adolescents with non-participating parents when they made their alcohol debut. They were also less likely to have ever been drunk in school year 9.</p> <p>Conclusion</p> <p>The results of the study suggested that Strong and Clear contributed to maintaining parents' restrictive attitude toward underage drinking during secondary school, postponing alcohol debut among the adolescents, and significantly reducing their drunkenness.</p

AKAP95 regulates splicing through scaffolding RNAs and RNA processing factors

YesAlternative splicing of pre-mRNAs significantly contributes to the complexity of gene expression in higher organisms, but the regulation of the splice site selection remains incompletely understood. We have previously demonstrated that a chromatin-associated protein, AKAP95 (AKAP8), has a remarkable activity in enhancing chromatin transcription. In this study, we have shown that AKAP95 physically interacts with many factors involved in transcription and RNA processing, and functionally regulates pre-mRNA splicing. AKAP95 directly promotes splicing in vitro and the inclusion of a specific exon of an endogenous gene FAM126A. The N-terminal YG-rich domain of AKAP95 is important for its binding to RNA processing factors including selective groups of hnRNP proteins, and its zinc finger domains are critical for pre-mRNA binding. Genome-wide binding assays revealed that AKAP95 bound preferentially to proximal intronic regions on a large number of pre-mRNAs in human transcriptome, and AKAP95 depletion predominantly resulted in reduced inclusion of many exons. AKAP95 also selectively coordinates with hnRNP H/F and U proteins in regulating alternative splicing events. We have further shown that AKAP95 directly interacts with itself. Taken together, our results establish AKAP95 as a novel and mostly positive regulator of premRNA splicing and a possible integrator of transcription and splicing regulation, and support a model that AKAP95 facilitates the splice site communication by looping out introns through both RNA-binding and protein-protein interaction.This work was supported by a UAB start-up fund to H.J

Relationship between Liver Mitochondrial Respiration and Proton Leak in Low and High RFI Steers from Two Lineages of RFI Angus Bulls.

The objective of this research is to evaluate liver mitochondrial oxygen consumption and proton leak kinetics in progeny from two lineages of Angus bulls with high and low residual feed intake (RFI). Two Angus bulls were selected based on results from a genetic test for RFI and were used as sires. Eight offspring at 10-11 months of age from each sire were housed in individual pens for 70-105 days following a diet adaptation period of 14 days. Progeny of the low RFI sire had 0.57 kg/d (P = 0.05) lower average RFI than progeny of the high RFI sire. There was no difference in dry matter intake between low and high RFI steers, but low RFI steers gained more body weight (P = 0.02) and tended to have higher average daily gains (P = 0.07). State 3 and State 4 respiration, RCR, and proton leak did not differ between high and low RFI steers (P = 0.96, P = 0.81, P = 0.93, and P = 0.88, resp.). Therefore, the increase in bodyweight gain which distinguished the low RFI steers from the high RFI steers may be associated with other metabolic mechanisms that are not associated with liver mitochondrial respiration and proton leak kinetics

Effect of motherhood on women's preferences for sources of health information: a prospective cohort study

To examine what sources of health information are preferred by first-time mothers-to-be and how these preferences change by the time their child reaches school age. Women expecting their first child (n = 649), recruited in a randomized trial of early childhood caries prevention at all five public maternity hospitals in Adelaide, were questioned about their preferences for health information. Their preferences were assessed again 4 and 7 years later. Answers at 7 years were compared with those of a population-based cohort of mothers with a first child of the same age. Parents were listed most frequently as a preferred source of health information during pregnancy (67.8%) followed by health care practitioners (48.8%). By the time the child reached school age, 78% listed health care practitioners as their preferred source compared with 15.5% listing parents, 21.7% friends and relatives, and 13% the Internet. Data from the population-based comparison group of mothers with a first child of similar age mimicked those of mothers enrolled in the trial. Mothers put a lot more trust in information received from health care professionals than they did before their child was born. This can create opportunities for enhancing the effectiveness of community health initiatives.Kamila Plutzer, Marc J. N. C. Keirs