Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques

Detalhes bibliográficos
Autor(a) principal: Monteiro, Ana Pereira de Brito Líbano
Data de Publicação: 2023
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10451/59248
Resumo: Tese de mestrado, Bioinformática e Biologia Computacional, 2023, Universidade de Lisboa, Faculdade de Ciências
id RCAP_c4d1a4c7d981937afbcbe2bf63b75b47
oai_identifier_str oai:repositorio.ul.pt:10451/59248
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniquesSplicing co-transcricionalmammalian native elongation transcript sequencing (mNET-seq)carboxyl terminal domain da polymerase II (Pol II CTD)inibição da transcriçãofosforilação da Tirosina 1Teses de mestrado - 2023Departamento de InformáticaTese de mestrado, Bioinformática e Biologia Computacional, 2023, Universidade de Lisboa, Faculdade de CiênciasMost RNA splicing – the excision of non-coding regions within precursor messenger RNA (mRNA) molecules – happens at the nascent moment, when the multiprotein complex RNA Polymerase II (Pol II) is synthesizing the new RNA molecules. Post-translational phosphorylations on the C-terminal domain (CTD) of the biggest Pol II subunit have been indicated as an inducer of different transcriptional and processivity moments. However, how a temporary inhibition of transcription influences these dynamics is still largely unexplored. In this study, a temporary transcription inhibition was promoted in Hela cells with the reversible drug 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB). To analyse co-transcriptional splicing, it was employed the mammalian native elongation transcript sequencing (mNET-seq) technique, that isolates the latest transcribed nucleotides from nascent pre-mRNAs, in a Pol II CTD-specific manner. The results of this work showed that there was a decrease of co-transcriptional splicing after treating cells with DRB. 47% of exons before DRB had splicing intermediates peaks – indicative of splicing first reaction – but only ~10% had these peaks 30 minutes after washing DRB. Splicing ratio distribution also shifted towards 0 after DRB treatment. It was also detected an increase of CTD Tyrosine 1 phosphorylation (Y1P) during and after the treatment. The absence of spliceosome components in Y1P elongation complex and the low levels of splicing signals in nascent RNA from Pol II Y1P suggest that this phosphorylation might not be associate with co-transcriptional splicing. This could explain the decrease observed after transcription restart. The association of CTD Serine 5 phosphorylation with splicing, discovered in previous studies, was also identified in these analyses, and barely affected by the DRB treatment. To conclude, these results suggest that higher Y1P levels inhibit co-transcription splicing, although further experiments are necessary to discriminate its effect from the influence of S5P.Fonseca, Maria CarmoCouto, Francisco José MoreiraRepositório da Universidade de LisboaMonteiro, Ana Pereira de Brito Líbano202320232026-02-13T00:00:00Z2023-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10451/59248enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T17:08:16Zoai:repositorio.ul.pt:10451/59248Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T22:09:11.970842Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
title Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
spellingShingle Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
Monteiro, Ana Pereira de Brito Líbano
Splicing co-transcricional
mammalian native elongation transcript sequencing (mNET-seq)
carboxyl terminal domain da polymerase II (Pol II CTD)
inibição da transcrição
fosforilação da Tirosina 1
Teses de mestrado - 2023
Departamento de Informática
title_short Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
title_full Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
title_fullStr Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
title_full_unstemmed Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
title_sort Unveiling co-transcriptional splicing kinetics using novel nascent transcriptomic techniques
author Monteiro, Ana Pereira de Brito Líbano
author_facet Monteiro, Ana Pereira de Brito Líbano
author_role author
dc.contributor.none.fl_str_mv Fonseca, Maria Carmo
Couto, Francisco José Moreira
Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Monteiro, Ana Pereira de Brito Líbano
dc.subject.por.fl_str_mv Splicing co-transcricional
mammalian native elongation transcript sequencing (mNET-seq)
carboxyl terminal domain da polymerase II (Pol II CTD)
inibição da transcrição
fosforilação da Tirosina 1
Teses de mestrado - 2023
Departamento de Informática
topic Splicing co-transcricional
mammalian native elongation transcript sequencing (mNET-seq)
carboxyl terminal domain da polymerase II (Pol II CTD)
inibição da transcrição
fosforilação da Tirosina 1
Teses de mestrado - 2023
Departamento de Informática
description Tese de mestrado, Bioinformática e Biologia Computacional, 2023, Universidade de Lisboa, Faculdade de Ciências
publishDate 2023
dc.date.none.fl_str_mv 2023
2023
2023-01-01T00:00:00Z
2026-02-13T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10451/59248
url http://hdl.handle.net/10451/59248
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/embargoedAccess
eu_rights_str_mv embargoedAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799134648173854720

Registros relacionados