{"id":{"repo_id":"cambridge","oai_identifier":"oai:www.repository.cam.ac.uk:1810/297676"},"canonical_url":"https://search.dev.ndltd.org/etd/cambridge/oai:www.repository.cam.ac.uk:1810/297676","repository":{"repo_id":"cambridge","name":"Cambridge University","base_url":"https://api.repository.cam.ac.uk/server/oai/request"},"display":{"title":"Identification of RBPMS as a smooth muscle master splicing regulator via association of its gene with super-enhancers","abstract":"Alternative splicing (AS) is primarily regulated by regulatory RNA-binding proteins (RBPs). It has been suggested that a small number of master splicing regulators might control cell-specific programs and these regulators could be identified via the association of their genes with transcriptional super-enhancers. Using this approach, RNA Binding Protein with Multiple Splicing (RBPMS) was identified as a critical splicing regulator in vascular smooth-muscle cells (SMCs). RBPMS is strongly downregulated during SMC dedifferentiation and is responsible for nearly 20% of the AS changes during this transition as indicated by mRNA-Seq of rat PAC1 cells with RBPMS-manipulated levels. RBPMS overexpression also promoted splicing events that are usually only observed in tissue SMCs. RBPMS targeted a network of proteins involved in the cytoskeleton and cell-adhesions, machineries remodelled during the transition from contractile to motile-dedifferentiated SMCs. RBPMS directly regulated target exons with a positional bias depending upon whether acting as an activator or repressor, as indicated by RBPMS-maps, in vivo transfections with minigene reporters, RBPMS RNA binding mutant, MS2 artificial tethering and lastly in vitro binding assays. RBPMS controlled splicing and activity of other splicing and post-transcriptional regulators (MBNL1, MBNL2 and LSM14B) as well as the key SMC transcription factor Myocardin. Structure-function analyses revealed that the two major RBPMS isoforms (RBPMS-A and B) have differential activity, and that dimerization and RBPMS C- terminus are essential to RBPMS splicing activity. Yet, RBPMS RRM was insufficient for splicing. In fact, a core section of the C-terminus of RBPMS-B antagonized its repressor-splicing activity. Additionally, two threonine residues of RBPMS could be phosphorylated differentially modulating RBPMS isoforms activity. Therefore, this study provides the strongest evidence to date for a molecular function of RBPMS as a splicing-regulator, matching many of the expected criteria of a master regulator of AS in differentiated VSMCs.","abstract_html":"Alternative splicing (AS) is primarily regulated by regulatory RNA-binding proteins (RBPs). It has been suggested that a small number of master splicing regulators might control cell-specific programs and these regulators could be identified via the association of their genes with transcriptional super-enhancers. Using this approach, RNA Binding Protein with Multiple Splicing (RBPMS) was identified as a critical splicing regulator in vascular smooth-muscle cells (SMCs). RBPMS is strongly downregulated during SMC dedifferentiation and is responsible for nearly 20% of the AS changes during this transition as indicated by mRNA-Seq of rat PAC1 cells with RBPMS-manipulated levels. RBPMS overexpression also promoted splicing events that are usually only observed in tissue SMCs. RBPMS targeted a network of proteins involved in the cytoskeleton and cell-adhesions, machineries remodelled during the transition from contractile to motile-dedifferentiated SMCs. RBPMS directly regulated target exons with a positional bias depending upon whether acting as an activator or repressor, as indicated by RBPMS-maps, in vivo transfections with minigene reporters, RBPMS RNA binding mutant, MS2 artificial tethering and lastly in vitro binding assays. RBPMS controlled splicing and activity of other splicing and post-transcriptional regulators (MBNL1, MBNL2 and LSM14B) as well as the key SMC transcription factor Myocardin. Structure-function analyses revealed that the two major RBPMS isoforms (RBPMS-A and B) have differential activity, and that dimerization and RBPMS C- terminus are essential to RBPMS splicing activity. Yet, RBPMS RRM was insufficient for splicing. In fact, a core section of the C-terminus of RBPMS-B antagonized its repressor-splicing activity. Additionally, two threonine residues of RBPMS could be phosphorylated differentially modulating RBPMS isoforms activity. Therefore, this study provides the strongest evidence to date for a molecular function of RBPMS as a splicing-regulator, matching many of the expected criteria of a master regulator of AS in differentiated VSMCs.","abstract_has_math":false,"creators":["Nakagaki Silva, Erick Eidy"],"institution":"University of Cambridge","degree_name":"Doctor of Philosophy (PhD)","degree_level":"Doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Smith, Christopher W J"],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-02-22","date_published":"2020-02-22","updated_at":"2026-07-22T22:24:01Z","subjects":["RBPMS","Vascular smooth muscle cells","Alternative splicing","PAC1 cells"],"languages":["en"],"rights":[],"rights_urls":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/6a874488-3313-47ef-bcb1-2616ddbfdd5b/download","https://www.rioxx.net/licenses/all-rights-reserved/"],"identifier_entries":[{"key":"dc:creator.authoridentifier","label":"Author Identifier","values":["0000000268784409"],"render_values":[{"text":"0000-0002-6878-4409","href":"https://orcid.org/0000-0002-6878-4409","code":true}]}]},"links":{"outbound_url":"https://doi.org/10.17863/CAM.44730","outbound_label":"DOI","outbound_source":"dc:identifier.doi"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Smith, Christopher W J"]},{"key":"dc:contributor.sponsor","label":"Sponsor","values":["British Heart Foundation (BHF) PG/16/28/32123); Wellcome 092900/Z/10/Z and 209368/Z/17/Z; MCTI | Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) 206813/2014-7"]},{"key":"dc:creator","label":"Author","values":["Nakagaki Silva, Erick Eidy"]},{"key":"dc:creator.authoridentifier","label":"Author Identifier","values":["0000000268784409"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2020-02-22"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Cambridge"]},{"key":"dc:relation.isreferencedby.uri","label":"Dc Relation Isreferencedby URI","values":["https://www.repository.cam.ac.uk/handle/1810/297676"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Doctoral"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["RBPMS","Vascular smooth muscle cells","Alternative splicing","PAC1 cells"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/6a874488-3313-47ef-bcb1-2616ddbfdd5b/download","https://www.rioxx.net/licenses/all-rights-reserved/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.doi","label":"DOI","values":["10.17863/CAM.44730"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/ca43f967-2c1f-4ae7-aa65-24108cb014d6/download"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Alternative splicing (AS) is primarily regulated by regulatory RNA-binding proteins (RBPs). It has been suggested that a small number of master splicing regulators might control cell-specific programs and these regulators could be identified via the association of their genes with transcriptional super-enhancers. Using this approach, RNA Binding Protein with Multiple Splicing (RBPMS) was identified as a critical splicing regulator in vascular smooth-muscle cells (SMCs). RBPMS is strongly downregulated during SMC dedifferentiation and is responsible for nearly 20% of the AS changes during this transition as indicated by mRNA-Seq of rat PAC1 cells with RBPMS-manipulated levels. RBPMS overexpression also promoted splicing events that are usually only observed in tissue SMCs. RBPMS targeted a network of proteins involved in the cytoskeleton and cell-adhesions, machineries remodelled during the transition from contractile to motile-dedifferentiated SMCs. RBPMS directly regulated target exons with a positional bias depending upon whether acting as an activator or repressor, as indicated by RBPMS-maps, in vivo transfections with minigene reporters, RBPMS RNA binding mutant, MS2 artificial tethering and lastly in vitro binding assays. RBPMS controlled splicing and activity of other splicing and post-transcriptional regulators (MBNL1, MBNL2 and LSM14B) as well as the key SMC transcription factor Myocardin. Structure-function analyses revealed that the two major RBPMS isoforms (RBPMS-A and B) have differential activity, and that dimerization and RBPMS C- terminus are essential to RBPMS splicing activity. Yet, RBPMS RRM was insufficient for splicing. In fact, a core section of the C-terminus of RBPMS-B antagonized its repressor-splicing activity. Additionally, two threonine residues of RBPMS could be phosphorylated differentially modulating RBPMS isoforms activity. Therefore, this study provides the strongest evidence to date for a molecular function of RBPMS as a splicing-regulator, matching many of the expected criteria of a master regulator of AS in differentiated VSMCs."]},{"key":"dc:format.checksum.md5","label":"Dc Format Checksum Md5","values":["115b2060ac51a2bf73ad165f0d2bd413","87eda9de84448d1f82354d60eee3eb5f"]},{"key":"dc:title","label":"Title","values":["Identification of RBPMS as a smooth muscle master splicing regulator via association of its gene with super-enhancers"]}]}],"canonical_facts":{"dc:contributor.advisor":["Smith, Christopher W J"],"dc:contributor.sponsor":["British Heart Foundation (BHF) PG/16/28/32123); Wellcome 092900/Z/10/Z and 209368/Z/17/Z; MCTI | Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) 206813/2014-7"],"dc:creator":["Nakagaki Silva, Erick Eidy"],"dc:creator.authoridentifier":["0000000268784409"],"dc:date.issued":["2020-02-22"],"dc:description.abstract":["Alternative splicing (AS) is primarily regulated by regulatory RNA-binding proteins (RBPs). It has been suggested that a small number of master splicing regulators might control cell-specific programs and these regulators could be identified via the association of their genes with transcriptional super-enhancers. Using this approach, RNA Binding Protein with Multiple Splicing (RBPMS) was identified as a critical splicing regulator in vascular smooth-muscle cells (SMCs). RBPMS is strongly downregulated during SMC dedifferentiation and is responsible for nearly 20% of the AS changes during this transition as indicated by mRNA-Seq of rat PAC1 cells with RBPMS-manipulated levels. RBPMS overexpression also promoted splicing events that are usually only observed in tissue SMCs. RBPMS targeted a network of proteins involved in the cytoskeleton and cell-adhesions, machineries remodelled during the transition from contractile to motile-dedifferentiated SMCs. RBPMS directly regulated target exons with a positional bias depending upon whether acting as an activator or repressor, as indicated by RBPMS-maps, in vivo transfections with minigene reporters, RBPMS RNA binding mutant, MS2 artificial tethering and lastly in vitro binding assays. RBPMS controlled splicing and activity of other splicing and post-transcriptional regulators (MBNL1, MBNL2 and LSM14B) as well as the key SMC transcription factor Myocardin. Structure-function analyses revealed that the two major RBPMS isoforms (RBPMS-A and B) have differential activity, and that dimerization and RBPMS C- terminus are essential to RBPMS splicing activity. Yet, RBPMS RRM was insufficient for splicing. In fact, a core section of the C-terminus of RBPMS-B antagonized its repressor-splicing activity. Additionally, two threonine residues of RBPMS could be phosphorylated differentially modulating RBPMS isoforms activity. Therefore, this study provides the strongest evidence to date for a molecular function of RBPMS as a splicing-regulator, matching many of the expected criteria of a master regulator of AS in differentiated VSMCs."],"dc:format.checksum.md5":["115b2060ac51a2bf73ad165f0d2bd413","87eda9de84448d1f82354d60eee3eb5f"],"dc:identifier.doi":["10.17863/CAM.44730"],"dc:identifier.uri":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/ca43f967-2c1f-4ae7-aa65-24108cb014d6/download"],"dc:language":["en"],"dc:publisher.institution":["University of Cambridge"],"dc:relation.isreferencedby.uri":["https://www.repository.cam.ac.uk/handle/1810/297676"],"dc:rights":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/6a874488-3313-47ef-bcb1-2616ddbfdd5b/download","https://www.rioxx.net/licenses/all-rights-reserved/"],"dc:subject":["RBPMS","Vascular smooth muscle cells","Alternative splicing","PAC1 cells"],"dc:title":["Identification of RBPMS as a smooth muscle master splicing regulator via association of its gene with super-enhancers"],"dc:type":["Thesis"],"dc:type.qualificationlevel":["Doctoral"],"dc:type.qualificationname":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-22T22:24:01Z"}