2 resultados para Post family

em Duke University


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Survival of a Perverse Nation traces the ways in which contemporary Armenian anxieties are congealing into the figure of the “homosexual.” As in other post-Soviet republics, homosexuality has increasingly become defined as the crisis of the times, and is understood by many as a destructive force linked to European encroachment. In Armenia, a growing right-wing nationalist movement since 2012 has been targeting LGBT and feminist activists. I suggest that this movement has arisen out of Armenia’s concerns regarding proper social and biological reproduction in the face of high rates of emigration of especially men in search of work. Many in the country blame this emigration on a post-Soviet oligarchy, with close ties to the government. This oligarchy, having quickly and massively privatized and liquidated industry and land during the war over the region of Nagorno-Karabagh (1990-1994) with Azerbaijan, created widespread un(der)employment. A national narrative attributing the nation’s survival of the 1915 Genocide and dispersion of its populations to strong morality preserved by institutions such as the Church and the family has now, in the post-Soviet era, ruptured into one of moral “perversion.” This dissertation is based on 15 months of ethnographic research, during which I participated in the work of two local non-governmental organizations: Public Information and Need for Knowledge, an LGBT rights organization and Women’s Resource Center, a feminist organization. I also conducted interviews with 150 households across Yerevan, the capital city, and did in-depth interviews with other activists, right-wing nationalists and journalists. Through psychoanalytic frameworks, as well as studies of kinship, I show how sovereignty – the longed for dream for Armenians over the last century – is felt to have failed because of the moral corruption of the illegitimate figures that fill Armenian seats of authority. I, thus, examine the ways in which a missing father of the household is discursively linked to the lack of strong leadership by a corrupt government, producing a prevalent feeling of moral disintegration that nationalists displace onto the “homosexual.”

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Post-transcriptional regulation of cytoplasmic mRNAs is an efficient mechanism of regulating the amounts of active protein within a eukaryotic cell. RNA sequence elements located in the untranslated regions of mRNAs can influence transcript degradation or translation through associations with RNA-binding proteins. Tristetraprolin (TTP) is the best known member of a family of CCCH zinc finger proteins that targets adenosine-uridine rich element (ARE) binding sites in the 3’ untranslated regions (UTRs) of mRNAs, promoting transcript deadenylation through the recruitment of deadenylases. More specifically, TTP has been shown to bind AREs located in the 3’-UTRs of transcripts with known roles in the inflammatory response. The mRNA-binding region of the protein is the highly conserved CCCH tandem zinc finger (TZF) domain. The synthetic TTP TZF domain has been shown to bind with high affinity to the 13-mer sequence of UUUUAUUUAUUUU. However, the binding affinities of full-length TTP family members to the same sequence and its variants are unknown. Furthermore, the distance needed between two overlapping or neighboring UUAUUUAUU 9-mers for tandem binding events of a full-length TTP family member to a target transcript has not been explored. To address these questions, we recombinantly expressed and purified the full-length C. albicans TTP family member Zfs1. Using full-length Zfs1, tagged at the N-terminus with maltose binding protein (MBP), we determined the binding affinities of the protein to the optimal TTP binding sequence, UUAUUUAUU. Fluorescence anisotropy experiments determined that the binding affinities of MBP-Zfs1 to non-canonical AREs were influenced by ionic buffer strength, suggesting that transcript selectivity may be affected by intracellular conditions. Furthermore, electrophoretic mobility shift assays (EMSAs) revealed that separation of two core AUUUA sequences by two uridines is sufficient for tandem binding of MBP-Zfs1. Finally, we found evidence for tandem binding of MBP-Zfs1 to a 27-base RNA oligonucleotide containing only a single ARE-binding site, and showed that this was concentration and RNA length dependent; this phenomenon had not been seen previously. These data suggest that the association of the TTP TZF domain and the TZF domains of other species, to ARE-binding sites is highly conserved. Domains outside of the TZF domain may mediate transcript selectivity in changing cellular conditions, and promote protein-RNA interactions not associated with the ARE-binding TZF domain.

In summary, the evidence presented here suggests that Zfs1-mediated decay of mRNA targets may require additional interactions, in addition to ARE-TZF domain associations, to promote transcript destabilization and degradation. These studies further our understanding of post-transcriptional steps in gene regulation.