Heterologous Expression of Immature Forms of Human Islet Amyloid Polypeptide in Yeast Triggers Intracellular Aggregation and Cytotoxicity

Detalhes bibliográficos
Autor(a) principal: Raimundo, Ana F.
Data de Publicação: 2020
Outros Autores: Ferreira, Sofia, Farrim, Maria I., Santos, Cláudia N., Menezes, Regina
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.
id RCAP_99a40b06298aa4dee014f939180f1e78
oai_identifier_str oai:run.unl.pt:10362/105562
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling 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
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_ 1799138019899342848

Registros relacionados