Post-Transcriptional Regulation of BCL2 mRNA by the RNA-Binding Protein ZFP36L1 in Malignant B Cells

The human ZFP36 zinc finger protein family consists of ZFP36, ZFP36L1, and ZFP36L2. These proteins regulate various cellular processes, including cell apoptosis, by binding to adenine uridine rich elements in the 3′ untranslated regions of sets of target mRNAs to promote their degradation. The pro-apoptotic and other functions of ZFP36 family members have been implicated in the pathogenesis of lymphoid malignancies. To identify candidate mRNAs that are targeted in the pro-apoptotic response by ZFP36L1, we reverse-engineered a gene regulatory network for all three ZFP36 family members using the ‘maximum information coefficient’ (MIC) for target gene inference on a large microarray gene expression dataset representing cells of diverse histological origin. Of the three inferred ZFP36L1 mRNA targets that were identified, we focussed on experimental validation of mRNA for the pro-survival protein, BCL2, as a target for ZFP36L1. RNA electrophoretic mobility shift assay experiments revealed that ZFP36L1 interacted with the BCL2 adenine uridine rich element. In murine BCL1 leukemia cells stably transduced with a ZFP36L1 ShRNA lentiviral construct, BCL2 mRNA degradation was significantly delayed compared to control lentiviral expressing cells and ZFP36L1 knockdown in different cell types (BCL1, ACHN, Ramos), resulted in increased levels of BCL2 mRNA levels compared to control cells. 3′ untranslated region luciferase reporter assays in HEK293T cells showed that wild type but not zinc finger mutant ZFP36L1 protein was able to downregulate a BCL2 construct containing the BCL2 adenine uridine rich element and removal of the adenine uridine rich core from the BCL2 3′ untranslated region in the reporter construct significantly reduced the ability of ZFP36L1 to mediate this effect. Taken together, our data are consistent with ZFP36L1 interacting with and mediating degradation of BCL2 mRNA as an important target through which ZFP36L1 mediates its pro-apoptotic effects in malignant B-cells.

The artist biopic: a historical analysis of narrative cinema, 1934-2010

The thesis provides an historical overview of the artist biopic that has emerged as a distinct sub-genre of the biopic as a whole, totalling some ninety films from Europe and America alone since the first talking artist biopic in 1934. Their making usually reflects a determination on the part of the director or star to see the artist as an alter-ego. Many of them were adaptations of successful literary works, which tempted financial backers by having a ready-made audience based on a pre-established reputation. The sub-genre’s development is explored via the grouping of films with associated themes and the use of case studies. These examples can then be used as models for exploring similar sets of data from other countries and time periods. The specific topics chosen for discussion include the representation of a single painter, for example, Vincent Van Gogh, to see how the treatment of an artist varies across several countries and over seventy years. British artist biopics are analysed as a case study in relation to the idea of them posing as a national stereotype. Topics within sex and gender studies are highlighted in analysis of the representation of the female artist and the queer artist as well as artists who have lived together as couples. A number of well-known gallery artists have become directors of artist biopics and their films are considered to see what particular insights a professional working artist can bring to the portrayal of artistic genius and creation. In the concluding part of the thesis it is argued that the artist biopic overall has survived the bad press which some individual productions have received and can even be said to have matured under the influence of directors producing a quality product for the art house, festival and avant-garde distribution circuits. As a genre it has proved extremely adaptable and has reflected the changing attitudes towards art and artists within the wider community. It has both encouraged renewed interest in the work of established national artists and also raised the profile of those relatively obscure such as Séraphine de Senlis and Pirosmani.