A Weakened Transcriptional Enhancer Yields Variegated Gene Expression

Identical genes in the same cellular environment are sometimes expressed differently. In some cases, including the immunoglobulin heavy chain (IgH) locus, this type of differential gene expression has been related to the absence of a transcriptional enhancer. To gain additional information on the role of the IgH enhancer, we examined expression driven by enhancers that were merely weakened, rather than fully deleted, using both mutations and insulators to impair enhancer activity. For this purpose we used a LoxP/Cre system to place a reporter gene at the same genomic site of a stable cell line. Whereas expression of the reporter gene was uniformly high in the presence of the normal, uninsulated enhancer and undetectable in its absence, weakened enhancers yielded variegated expression of the reporter gene; i.e., the average level of expression of the same gene differed in different clones, and expression varied significantly among cells within individual clones. These results indicate that the weakened enhancer allows the reporter gene to exist in at least two states. Subtle aspects of the variegation suggest that the IgH enhancer decreases the average duration (half-life) of the silent state. This analysis has also tested the conventional wisdom that enhancer activity is independent of distance and orientation. Thus, our analysis of mutant (truncated) forms of the IgH enhancer revealed that the 250 bp core enhancer was active in its normal position, ∼1.4 kb 3′ of the promoter, but inactive ∼6 kb 3′, indicating that the activity of the core enhancer was distance-dependent. A longer segment – the core enhancer plus ∼1 kb of 3′ flanking material, including the 3′ matrix attachment region – was active, and the activity of this longer segment was orientation-dependent. Our data suggest that this 3′ flank includes binding sites for at least two activators

Mixed marriages and transnational families in the intercultural context : a case study of African-Spanish couples in Catalonia, Spain

Premi a l'excel·lència investigadora. Àmbit de les Ciències Socials. 2008One of the consequences of international migration and the permanent settlement of immigrants in southern EU countries is the growing number of inter-country marriages and the formation of transnational families. Using both quantitative and qualitative data, this article examines patterns of endogamy and exogamy (i.e. marriage within/outside a particular group or category) among African immigrants in Catalonia, focusing on bi-national Senegalese- and Gambian-Spanish couples. Socio-demographic profiles, transnationality, the dynamics of cultural change or retention, and the formation of transcultural identities are explored. The evidence presented suggests that social-class factors are more important than cultural origins in patterns of endogamy and exogamy, in the dynamics of living together and in the bringing-up of children of mixed unions. Such a conclusion negates culturalists' explanations of endogamy and exogamy while, at the same time, emphasising the role of social actors as active subjects in these processes. I further argue that mixed couples and their offspring deal-to a greater or lesser extent-with multiple localisations and cultural backgrounds (i.e. here and there), rather than experiencing a 'clash between two cultures'. Therefore, it would be a mistake to pretend that multicultural links do not exist and that they cannot be revitalised and functional. The paper starts and ends by addressing the complexities of processes of interculturalism, resisting an interpretation of hybridity and segregation as contradictory or exclusive realities

Financial journalism, conflicts of interest and ethics: a case study of Hong Kong

This article explores the practice, ethics, and regulation of financial and business journalism in Hong Kong and examines business journalists' understanding of their “social responsibility.” The research is based on an analysis of the legal framework and codes of conduct plus interviews with professional journalists, editors, and other experts. The focus is on the interrelated issues of conflicts of interest, disclosure of interest, investment by journalists, and market manipulation. While some journalists are aware of key ethical dilemmas and the professional standards, there remains confusion among many journalists regarding appropriate standards. The Chinese language business media in Hong Kong operate with a more relaxed approach to conflicts of interest than the English language media and global business news providers. The assumptions behind these different approaches are compared and contrasted

A functional screen identifies hDRIL1 as an oncogene that rescues RAS-induced senescence

Primary fibroblasts respond to activated H-RASV12 by undergoing premature arrest, which resembles replicative senescence1. This irreversible 'fail-safe mechanism' requires p19ARF, p53 and the Retinoblastoma (Rb) family: upon their disruption, RASV12-expressing cells fail to undergo senescence and continue to proliferate1, 2, 3, 4, 5, 6, 7. Similarly, co-expression of oncogenes such as c-MYC or E1A rescues RASV12-induced senescence. To identify novel genes that allow escape from RASV12-induced senescence, we designed an unbiased, retroviral complementary DNA library screen. We report on the identification of DRIL1, the human orthologue of the mouse Bright and Drosophila dead ringer transcriptional regulators. DRIL1 renders primary murine fibroblasts unresponsive to RASV12-induced anti-proliferative signalling by p19ARF/p53/p21CIP1, as well as by p16INK4a. In this way, DRIL1 not only rescues RASV12-induced senescence but also causes these fibroblasts to become highly oncogenic. Furthermore, DRIL1 immortalizes mouse fibroblasts, in the presence of high levels of p16INK4a. Immortalization by DRIL1, whose product binds the pRB-controlled transcription factor E2F1 (ref. 8), is correlated with induction of E2F1 activity. Correspondingly, DRIL1 induces the E2F1 target Cyclin E1, overexpression of which is sufficient to trigger escape from senescence. Thus, DRIL1 disrupts cellular protection against RASV12-induced proliferation downstream of the p19ARF/p53 pathway

Dorsal-Mediated Repression Requires the Formation of a Multiprotein Repression Complex at the Ventral Silencer

Dorsal functions as both an activator and repressor of transcription to determine dorsoventral fate in the Drosophila melanogaster embryo. Repression by Dorsal requires the corepressor Groucho (Gro) and is mediated by silencers termed ventral repression regions (VRRs). A VRR in zerknüllt (zen) contains Dorsal binding sites as well as an essential element termed AT2. We have identified and purified an AT2 DNA binding activity in embryos and shown it to consist of cut (ct) and dead ringer (dri) gene products. Studies of loss-of-function mutations in ct and dri demonstrate that both genes are required for the activity of the AT2 site. Dorsal and Dri both bind Gro, acting cooperatively to recruit it to the DNA. Thus, ventral repression may require the formation of a multiprotein complex at the VRR. This complex includes Dorsal, Gro, and additional DNA binding proteins, which appear to convert Dorsal from an activator to a repressor by enabling it to recruit Gro to the template. By showing how binding site context can dramatically alter transcription factor function, these findings help clarify the mechanisms responsible for the regulatory specificity of transcription factors