{"id":{"repo_id":"utswmed","oai_identifier":"oai:utswmed-ir.tdl.org:2152.5/10803"},"canonical_url":"https://search.dev.ndltd.org/etd/utswmed/oai:utswmed-ir.tdl.org:2152.5/10803","repository":{"repo_id":"utswmed","name":"University of Texas Southwestern Medical Center","base_url":"https://utswmed-ir.tdl.org/server/oai/request"},"display":{"title":"Functional Analysis of KAP1 Requirements for HIV-1 Transcription and Latency Reactivation","abstract":"HIV-1 latency maintenance and reactivation are regulated by several viral and host factors. One such regulator is Krüppel-associated box (KRAB)-associated protein 1 (KAP1: also named TRIM28 or TIF1β). While initial studies have revealed KAP1 to be a positive regulator of latency reversal in transformed and primary CD4+ T cells, subsequent studies have proposed KAP1 to be a repressor required for latency maintenance. Given this discrepancy, here we re-examine KAP1 transcription regulatory functions using a chemical genetics strategy to acutely deplete KAP1 expression to avoid the accumulation of indirect effects. KAP1 acute loss dampened HIV-1 promoter activity in response to activating signals, a function that can be restored upon complementation with exogenous KAP1, thus revealing KAP1-mediated activation is on target. By combining comprehensive KAP1 domain deletion and mutagenesis in a cell-based reporter assay, I genetically defined the RING finger domain and an Intrinsically Disordered Region as key activating features. However, KAP1 acute loss in a physiological relevant model, reveals limitations compared to chronic silencing of KAP1 in the same system. I also identified a potential co-factor MAGE-D2 from previous biochemical and genetic screens that interacts with KAP1. MAGE-D2 mimics KAP1s ability to activate HIV-1 latency reactivation in response to stimulation suggesting they form a complex to regulate the proviral genome. Together, this study solidifies the notion that KAP1 activates HIV-1 transcription by exploiting its multi-domain protein arrangement through previously unknown domains and functions. Also, this study highlights the need for better techniques to assess the direct consequence of KAP1 loss.","abstract_html":"HIV-1 latency maintenance and reactivation are regulated by several viral and host factors. One such regulator is Krüppel-associated box (KRAB)-associated protein 1 (KAP1: also named TRIM28 or TIF1β). While initial studies have revealed KAP1 to be a positive regulator of latency reversal in transformed and primary CD4+ T cells, subsequent studies have proposed KAP1 to be a repressor required for latency maintenance. Given this discrepancy, here we re-examine KAP1 transcription regulatory functions using a chemical genetics strategy to acutely deplete KAP1 expression to avoid the accumulation of indirect effects. KAP1 acute loss dampened HIV-1 promoter activity in response to activating signals, a function that can be restored upon complementation with exogenous KAP1, thus revealing KAP1-mediated activation is on target. By combining comprehensive KAP1 domain deletion and mutagenesis in a cell-based reporter assay, I genetically defined the RING finger domain and an Intrinsically Disordered Region as key activating features. However, KAP1 acute loss in a physiological relevant model, reveals limitations compared to chronic silencing of KAP1 in the same system. I also identified a potential co-factor MAGE-D2 from previous biochemical and genetic screens that interacts with KAP1. MAGE-D2 mimics KAP1s ability to activate HIV-1 latency reactivation in response to stimulation suggesting they form a complex to regulate the proviral genome. Together, this study solidifies the notion that KAP1 activates HIV-1 transcription by exploiting its multi-domain protein arrangement through previously unknown domains and functions. Also, this study highlights the need for better techniques to assess the direct consequence of KAP1 loss.","abstract_has_math":false,"creators":["Randolph, Keyera"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Conrad, Nicholas","Reese, Tiffany A.","Nijhawan, Deepak","D'Orso, Iván"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2026,"date_issued":"2026-06-15T19:42:14Z","date_published":"2026-06-15T19:42:14Z","updated_at":"2026-07-24T05:52:15Z","subjects":["HIV-1","Transcriptional Activation","Tripartite Motif-Containing Protein 28"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["1596185286"],"render_values":[{"text":"1596185286","href":null,"code":true}]}]},"links":{"outbound_url":"https://hdl.handle.net/2152.5/10803","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Conrad, Nicholas","Reese, Tiffany A.","Nijhawan, Deepak","D'Orso, Iván"]},{"key":"dc:creator","label":"Author","values":["Randolph, Keyera"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2026-06-15T19:42:14Z","2024-05","May 2024","2026-06-15T19:42:15Z"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","text"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["HIV-1","Transcriptional Activation","Tripartite Motif-Containing Protein 28"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://hdl.handle.net/2152.5/10803","1596185286"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["HIV-1 latency maintenance and reactivation are regulated by several viral and host factors. One such regulator is Krüppel-associated box (KRAB)-associated protein 1 (KAP1: also named TRIM28 or TIF1β). While initial studies have revealed KAP1 to be a positive regulator of latency reversal in transformed and primary CD4+ T cells, subsequent studies have proposed KAP1 to be a repressor required for latency maintenance. Given this discrepancy, here we re-examine KAP1 transcription regulatory functions using a chemical genetics strategy to acutely deplete KAP1 expression to avoid the accumulation of indirect effects. KAP1 acute loss dampened HIV-1 promoter activity in response to activating signals, a function that can be restored upon complementation with exogenous KAP1, thus revealing KAP1-mediated activation is on target. By combining comprehensive KAP1 domain deletion and mutagenesis in a cell-based reporter assay, I genetically defined the RING finger domain and an Intrinsically Disordered Region as key activating features. However, KAP1 acute loss in a physiological relevant model, reveals limitations compared to chronic silencing of KAP1 in the same system. I also identified a potential co-factor MAGE-D2 from previous biochemical and genetic screens that interacts with KAP1. MAGE-D2 mimics KAP1s ability to activate HIV-1 latency reactivation in response to stimulation suggesting they form a complex to regulate the proviral genome. Together, this study solidifies the notion that KAP1 activates HIV-1 transcription by exploiting its multi-domain protein arrangement through previously unknown domains and functions. Also, this study highlights the need for better techniques to assess the direct consequence of KAP1 loss."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Functional Analysis of KAP1 Requirements for HIV-1 Transcription and Latency Reactivation"]}]}],"canonical_facts":{"dc:contributor":["Conrad, Nicholas","Reese, Tiffany A.","Nijhawan, Deepak","D'Orso, Iván"],"dc:creator":["Randolph, Keyera"],"dc:date":["2026-06-15T19:42:14Z","2024-05","May 2024","2026-06-15T19:42:15Z"],"dc:description":["HIV-1 latency maintenance and reactivation are regulated by several viral and host factors. One such regulator is Krüppel-associated box (KRAB)-associated protein 1 (KAP1: also named TRIM28 or TIF1β). While initial studies have revealed KAP1 to be a positive regulator of latency reversal in transformed and primary CD4+ T cells, subsequent studies have proposed KAP1 to be a repressor required for latency maintenance. Given this discrepancy, here we re-examine KAP1 transcription regulatory functions using a chemical genetics strategy to acutely deplete KAP1 expression to avoid the accumulation of indirect effects. KAP1 acute loss dampened HIV-1 promoter activity in response to activating signals, a function that can be restored upon complementation with exogenous KAP1, thus revealing KAP1-mediated activation is on target. By combining comprehensive KAP1 domain deletion and mutagenesis in a cell-based reporter assay, I genetically defined the RING finger domain and an Intrinsically Disordered Region as key activating features. However, KAP1 acute loss in a physiological relevant model, reveals limitations compared to chronic silencing of KAP1 in the same system. I also identified a potential co-factor MAGE-D2 from previous biochemical and genetic screens that interacts with KAP1. MAGE-D2 mimics KAP1s ability to activate HIV-1 latency reactivation in response to stimulation suggesting they form a complex to regulate the proviral genome. Together, this study solidifies the notion that KAP1 activates HIV-1 transcription by exploiting its multi-domain protein arrangement through previously unknown domains and functions. Also, this study highlights the need for better techniques to assess the direct consequence of KAP1 loss."],"dc:format":["application/pdf"],"dc:identifier":["https://hdl.handle.net/2152.5/10803","1596185286"],"dc:language":["en"],"dc:subject":["HIV-1","Transcriptional Activation","Tripartite Motif-Containing Protein 28"],"dc:title":["Functional Analysis of KAP1 Requirements for HIV-1 Transcription and Latency Reactivation"],"dc:type":["Thesis","text"]},"updated_at":"2026-07-24T05:52:15Z"}