Type I Hsp40s/DnaJs aggregates exhibit features reminiscent of amyloidogenic structures

Autor(es)

Tiroli-Cepeda, Ana O. ; Linhares, Leonardo A. ; Aragao, Annelize Z. B. ; de Jesus, Jemmyson R. ; Wasilewska-Sampaio, Ana P. ; De Felice, Fernanda G. ; Ferreira, Sergio T. ; Borges, Julio C. ; Cyr, Douglas M. ; Ramos, Carlos H. I.

• Tipo de documento: Artigo Científico
• Fonte: FEBS Journal; v. 291, n. 17, p. 20-pg., 2024-07-08.

Resumo

A rise in temperature triggers a structural change in the human Type I 40 kDa heat shock protein (Hsp40/DnaJ), known as DNAJA1. This change leads to a less compact structure, characterized by an increased presence of solvent-exposed hydrophobic patches and beta-sheet-rich regions. This transformation is validated by circular dichroism, thioflavin T binding, and Bis-ANS assays. The formation of this beta-sheet-rich conformation, which is amplified in the absence of zinc, leads to protein aggregation. This aggregation is induced not only by high temperatures but also by low ionic strength and high protein concentration. The aggregated conformation exhibits characteristics of an amyloidogenic structure, including a distinctive X-ray diffraction pattern, seeding competence (which stimulates the formation of amyloid-like aggregates), cytotoxicity, resistance to SDS, and fibril formation. Interestingly, the yeast Type I Ydj1 also tends to adopt a similar beta-sheet-rich structure under comparable conditions, whereas Type II Hsp40s, whether human or from yeast, do not. Moreover, Ydj1 aggregates were found to be cytotoxic. Studies using DNAJA1- and Ydj1-deleted mutants suggest that the zinc-finger region plays a crucial role in amyloid formation. Our discovery of amyloid aggregation in a C-terminal deletion mutant of DNAJA1, which resembles a spliced homolog expressed in the testis, implies that Type I Hsp40 co-chaperones may generate amyloidogenic species in vivo. (AU)

• Processo FAPESP: 07/04529-5 - Estudo do mecanismo de interacao entre hsp70 e chaperones com funcao de reconhecimento e apresentacao de substratos.
• Beneficiário: Ana Olivia Tiroli Cepeda
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 18/00768-0 - Correlação entre metais e homeostase proteica: estudos iniciais investigando a incorporação do isótopo 68Zn na co-chaperona Hsp40 e seu efeito em levedura
• Beneficiário: Jemmyson Romário de Jesus
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/26131-5 - Chaperoma: estudo da relação entre a estrutura dos seus componentes e a manutenção da proteostase
• Beneficiário: Carlos Henrique Inacio Ramos
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/11986-5 - Geração e Armazenamento de Novas Energias: trazendo desenvolvimento tecnológico para o país
• Beneficiário: Ana Flávia Nogueira
• Modalidade de apoio: Auxílio à Pesquisa - Programa Centros de Pesquisa em Engenharia
• Processo FAPESP: 12/50161-8 - Estudo da estrutura e função da chaperona Hsp90 com ênfase no seu papel em homeostase celular
• Beneficiário: Carlos Henrique Inacio Ramos
• Modalidade de apoio: Auxílio à Pesquisa - Temático