Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/105562 |
Resumo: | Funding: iNOVA4Health – UID/Multi/04462/2019, a program financially supported by Fundação para a Ciência e Tecnologia/Ministério da Educação e Ciência, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement is acknowledged. Funding from INTERFACE Programme, through the Innovation, Technology and Circular Economy Fund (FITEC), is gratefully acknowledged. This study was also supported by FCT via PTDC/BIA-MOL31104/2017, UID/Multi/04462/2013- SubProj iNOVA4Health 44, and UID/Multi/04462/2019-SubProj iNOVA4Health C44 to RM, PD/BD/135504/2018 to AR. Sociedade Portuguesa de Diabetologia for the Nuno CasteloBranco Prize – 2016 attributed to RM is also acknowledged. |
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7160 |
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Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicityimmature islet amyloid polypeptideislet amyloid polypeptideislet amyloid polypeptide-induced toxicityoligomerizationprotein aggregationyeast modelMicrobiologyMicrobiology (medical)SDG 3 - Good Health and Well-beingFunding: iNOVA4Health – UID/Multi/04462/2019, a program financially supported by Fundação para a Ciência e Tecnologia/Ministério da Educação e Ciência, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement is acknowledged. Funding from INTERFACE Programme, through the Innovation, Technology and Circular Economy Fund (FITEC), is gratefully acknowledged. This study was also supported by FCT via PTDC/BIA-MOL31104/2017, UID/Multi/04462/2013- SubProj iNOVA4Health 44, and UID/Multi/04462/2019-SubProj iNOVA4Health C44 to RM, PD/BD/135504/2018 to AR. Sociedade Portuguesa de Diabetologia for the Nuno CasteloBranco Prize – 2016 attributed to RM is also acknowledged.Diabetes is a major public health issue that has attained alarming levels worldwide. Pancreatic aggregates of human islet amyloid polypeptide (IAPP) represent a major histopathological hallmark of type 2 diabetes. IAPP is expressed in β-cells as pre-pro-IAPP (ppIAPP) that is first processed to pro-IAPP (pIAPP) and finally to its mature form (matIAPP), being released upon glucose stimulation together with insulin. Impairment and overload of the IAPP processing machinery seem to be associated with the accumulation of immature IAPP species and the formation of toxic intracellular oligomers, which have been associated with β-cell dyshomeostasis and apoptosis. Nevertheless, the pathological importance of these immature IAPP forms for the assembly and cytotoxicity of these oligomers is not completely understood. Here, we describe the generation and characterization of unprecedented Saccharomyces cerevisiae models recapitulating IAPP intracellular oligomerization. Expression of green fluorescent protein (GFP) fusions of human ppIAPP, pIAPP, and matIAPP proved to be toxic in yeast cells at different extents, with ppIAPP exerting the most deleterious effect on yeast growth and cell viability. Although expression of all IAPP constructs induced the formation of intracellular aggregates in yeast cells, our data point out the accumulation of insoluble oligomeric species enriched in immature ppIAPP as the trigger of the high toxicity mediated by this construct in cells expressing ppIAPP-GFP. In addition, MS/MS analysis indicated that oligomeric species found in the ppIAPP-GFP lysates contain the N-terminal sequence of the propeptide fused to GFP. These models represent powerful tools for future research focused on the relevance of immature forms in IAPP-induced toxicity. Furthermore, they are extremely useful in high-throughput screenings for genetic and chemical modulators of IAPP aggregation.Centro de Estudos de Doenças Crónicas (CEDOC)Instituto de Tecnologia Química e Biológica António Xavier (ITQB)NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)RUNRaimundo, Ana F.Ferreira, SofiaFarrim, Maria I.Santos, Cláudia N.Menezes, Regina2020-10-13T22:43:57Z2020-09-032020-09-03T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/105562eng1664-302XPURE: 20226289https://doi.org/10.3389/fmicb.2020.02035info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T04:50:46Zoai:run.unl.pt:10362/105562Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:40:31.033829Repositório 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 |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
title |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
spellingShingle |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity Raimundo, Ana F. immature islet amyloid polypeptide islet amyloid polypeptide islet amyloid polypeptide-induced toxicity oligomerization protein aggregation yeast model Microbiology Microbiology (medical) SDG 3 - Good Health and Well-being |
title_short |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
title_full |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
title_fullStr |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
title_full_unstemmed |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
title_sort |
Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity |
author |
Raimundo, Ana F. |
author_facet |
Raimundo, Ana F. Ferreira, Sofia Farrim, Maria I. Santos, Cláudia N. Menezes, Regina |
author_role |
author |
author2 |
Ferreira, Sofia Farrim, Maria I. Santos, Cláudia N. Menezes, Regina |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Centro de Estudos de Doenças Crónicas (CEDOC) Instituto de Tecnologia Química e Biológica António Xavier (ITQB) NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM) RUN |
dc.contributor.author.fl_str_mv |
Raimundo, Ana F. Ferreira, Sofia Farrim, Maria I. Santos, Cláudia N. Menezes, Regina |
dc.subject.por.fl_str_mv |
immature islet amyloid polypeptide islet amyloid polypeptide islet amyloid polypeptide-induced toxicity oligomerization protein aggregation yeast model Microbiology Microbiology (medical) SDG 3 - Good Health and Well-being |
topic |
immature islet amyloid polypeptide islet amyloid polypeptide islet amyloid polypeptide-induced toxicity oligomerization protein aggregation yeast model Microbiology Microbiology (medical) SDG 3 - Good Health and Well-being |
description |
Funding: iNOVA4Health – UID/Multi/04462/2019, a program financially supported by Fundação para a Ciência e Tecnologia/Ministério da Educação e Ciência, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement is acknowledged. Funding from INTERFACE Programme, through the Innovation, Technology and Circular Economy Fund (FITEC), is gratefully acknowledged. This study was also supported by FCT via PTDC/BIA-MOL31104/2017, UID/Multi/04462/2013- SubProj iNOVA4Health 44, and UID/Multi/04462/2019-SubProj iNOVA4Health C44 to RM, PD/BD/135504/2018 to AR. Sociedade Portuguesa de Diabetologia for the Nuno CasteloBranco Prize – 2016 attributed to RM is also acknowledged. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-10-13T22:43:57Z 2020-09-03 2020-09-03T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/105562 |
url |
http://hdl.handle.net/10362/105562 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1664-302X PURE: 20226289 https://doi.org/10.3389/fmicb.2020.02035 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
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 |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799138019899342848 |