Inclusive Scholarship: Developing Black Studies in the United States

Brings together four reports commissioned between 1982 and 2000 that examine the history of African American Studies, its impact, and its institutionalization. Reviews Ford's grantmaking to African American Studies programs from 1982 to 2007

Definition of Naturally Processed Peptides Reveals Convergent Presentation of Autoantigenic Topoisomerase I Epitopes in Scleroderma.

ObjectiveAutoimmune responses to DNA topoisomerase I (topo I) are found in a subset of scleroderma patients who are at high risk for interstitial lung disease (ILD) and mortality. Anti-topo I antibodies (ATAs) are associated with specific HLA-DRB1 alleles, and the frequency of HLA-DR-restricted topo I-specific CD4+ T cells is associated with the presence, severity, and progression of ILD. Although this strongly implicates the presentation of topo I peptides by HLA-DR in scleroderma pathogenesis, the processing and presentation of topo I has not been studied.MethodsWe developed a natural antigen processing assay (NAPA) to identify putative CD4+ T cell epitopes of topo I presented by monocyte-derived dendritic cells (mo-DCs) from 6 ATA-positive patients with scleroderma. Mo-DCs were pulsed with topo I protein, HLA-DR-peptide complexes were isolated, and eluted peptides were analyzed by mass spectrometry. We then examined the ability of these naturally presented peptides to induce CD4+ T cell activation in 11 ATA-positive and 11 ATA-negative scleroderma patients.ResultsWe found that a common set of 10 topo I epitopes was presented by Mo-DCs from scleroderma patients with diverse HLA-DR variants. Sequence analysis revealed shared peptide-binding motifs within the HLA-DRβ chains of ATA-positive patients and a subset of topo I epitopes with distinct sets of anchor residues capable of binding to multiple different HLA-DR variants. The NAPA-derived epitopes elicited robust CD4+ T cell responses in 73% of ATA-positive patients (8 of 11), and the number of epitopes recognized correlated with ILD severity (P = 0.025).ConclusionThese findings mechanistically implicate the presentation of a convergent set of topo I epitopes in the development of scleroderma

The jazz singer's legacy: The racial role-play of African-Americans and Jews in twentieth-century American performance.

This study re-examines the racial role-play of African-Americans and Jewish-Americans to further understand how racial identity has been constructed in America and the ways in which these constructions overlap and affect one another. Although racial role-play is most often associated with blackface minstrelsy, I define it more broadly as an overt staging of a racial identity. Unlike passing---an attempt to assume another identity without being detected---racial role-play calls attention to its performance. Because of its negative association with blackface minstrelsy, racial role-play disturbs many Americans. Yet to dismiss or avoid the function racial role-play has had in America is to miss a key means of understanding the cultural formation of our country. Moreover, racial role-play has been used by performers of various backgrounds, including African-Americans, to undermine stereotypes, to cross racial boundaries, and to question racial definitions altogether.Chapter One discusses the history of minstrelsy and how this practice might have been used and viewed by both African-Americans and Jewish-Americans. Chapter Two follows this discussion with an in-depth look at both the play and the film of The Jazz Singer, including critical responses from the Jewish, the Black, and the mainstream press. In contrast to Whiteness scholars who argue that blackface was a means for Jews to distance themselves from Blacks in order to assimilate into White America, I argue that blackface was a form of identification with African-Americans in order to distinguish themselves from White Christians. Chapter Three focuses on the attempts of certain African-American artists in the 1960s to re-appropriate racial role-play and demonstrates how Jews functioned symbolically as a means of asserting an evolving Black identity. Chapter Four illustrates how Jews once again utilized racial role-play alongside of African-Americans in the 1990s as performance artists explored identity politics. In particular, I examine the work of Anna Deavere Smith, Sandra Bernhard, and Danny Hoch---who suggest that identities are not fixed, but rather can be continually questioned, challenged, and negotiated through performance. This fluidity, however, comes at the cost of losing or ignoring group distinctions, whether these distinctions are self-imposed or externally assigned.Thesis (Ph.D.)--Columbia University, 2004.School code: 0054

The cardiac acetyl-lysine proteome.

In the heart, lysine acetylation has been implicated in processes ranging from transcriptional control of pathological remodeling, to cardioprotection arising from caloric restriction. Given the emerging importance of this post-translational modification, we used a proteomic approach to investigate the broader role of lysine acetylation in the heart using a guinea pig model. Briefly, hearts were fractionated into myofilament-, mitochondrial- and cytosol-enriched fractions prior to proteolysis and affinity-enrichment of acetylated peptides. LC-MS/MS analysis identified 1075 acetylated peptides, harboring 994 acetylation sites that map to 240 proteins with a global protein false discovery rate <0.8%. Mitochondrial targets account for 59% of identified proteins and 64% of sites. The majority of the acetyl-proteins are enzymes involved in fatty acid metabolism, oxidative phosphorylation or the TCA cycle. Within the cytosolic fraction, the enzymes of glycolysis, fatty acid synthesis and lipid binding are prominent. Nuclear targets included histones and the transcriptional regulators E1A(p300) and CREB binding protein. Comparison of our dataset with three previous global acetylomic studies uniquely revealed 53 lysine-acetylated proteins. Specifically, newly-identified acetyl-proteins include Ca(2+)-handling proteins, RyR2 and SERCA2, and the myofilament proteins, myosin heavy chain, myosin light chains and subunits of the Troponin complex, among others. These observations were confirmed by anti-acetyl-lysine immunoblotting. In summary, cardiac lysine acetylation may play a role in cardiac substrate selection, bioenergetic performance, and maintenance of redox balance. New sites suggest a host of potential mechanisms by which excitation-contraction coupling may also be modulated

Phosphorylation of mutant huntingtin at serine 116 modulates neuronal toxicity.

Phosphorylation has been shown to have a significant impact on expanded huntingtin-mediated cellular toxicity. Several phosphorylation sites have been identified on the huntingtin (Htt) protein. To find new potential therapeutic targets for Huntington's Disease (HD), we used mass spectrometry to identify novel phosphorylation sites on N-terminal Htt, expressed in HEK293 cells. Using site-directed mutagenesis we introduced alterations of phosphorylation sites in a N586 Htt construct containing 82 polyglutamine repeats. The effects of these alterations on expanded Htt toxicity were evaluated in primary neurons using a nuclear condensation assay and a direct time-lapse imaging of neuronal death. As a result of these studies, we identified several novel phosphorylation sites, validated several known sites, and discovered one phospho-null alteration, S116A, that had a protective effect against expanded polyglutamine-mediated cellular toxicity. The results suggest that S116 is a potential therapeutic target, and indicate that our screening method is useful for identifying candidate phosphorylation sites

Combined antibody/lectin enrichment identifies extensive changes in the O-GlcNAc sub-proteome upon oxidative stress

O-Linked N-acetyl-β-d-glucosamine (O-GlcNAc) is a dynamic post-translational modification that modifies and regulates over 3000 nuclear, cytoplasmic, and mitochondrial proteins. Upon exposure to stress and injury, cells and tissues increase the O-GlcNAc modification, or O-GlcNAcylation, of numerous proteins promoting the cellular stress response and thus survival. The aim of this study was to identify proteins that are differentially O-GlcNAcylated upon acute oxidative stress (H₂O₂) to provide insight into the mechanisms by which O-GlcNAc promotes survival. We achieved this goal by employing Stable Isotope Labeling of Amino Acids in Cell Culture (SILAC) and a novel "G5-lectibody" immunoprecipitation strategy that combines four O-GlcNAc-specific antibodies (CTD110.6, RL2, HGAC39, and HGAC85) and the lectin WGA. Using the G5-lectibody column in combination with basic reversed phase chromatography and C₁₈ RPLC-MS/MS, 990 proteins were identified and quantified. Hundreds of proteins that were identified demonstrated increased (>250) or decreased (>110) association with the G5-lectibody column upon oxidative stress, of which we validated the O-GlcNAcylation status of 24 proteins. Analysis of proteins with altered glycosylation suggests that stress-induced changes in O-GlcNAcylation cluster into pathways known to regulate the cell's response to injury and include protein folding, transcriptional regulation, epigenetics, and proteins involved in RNA biogenesis. Together, these data suggest that stress-induced O-GlcNAcylation regulates numerous and diverse cellular pathways to promote cell and tissue survival.19 page(s

Generation and expression of N586-82Q constructs with phospho-site alterations.

<p>(A) Schematic representation of the Htt N586 and the sites altered. Italics indicate previously described phosphorylation sites. Detected sites are in bold. (B) Expression of constructs in primary cortical neurons. (C) Quantification of expression of N586 constructs in primary neurons. Fluorescence was quantified using Volocity, and results are expressed as percent of N586-22Q level of expression. (D) Western blot of expression of N586 constructs with phospho-site alterations in primary cortical neurons. The image shown is representative of 4 replicate experiments.</p