In vivo observation of amyloid-like fibrils produced under stress

Author(s)

Fontana, Natalia A. ; Rosse, Ariane D. ; Watts, Anthony ; Coelho, Paulo S. R. ; Costa-Filho, Antonio J.

• Document type: Journal article
• Source: International Journal of Biological Macromolecules; v. 199, p. 9-pg., 2022-01-04.

Abstract

The participation of amyloids in neurodegenerative diseases and functional processes has triggered the quest for methods allowing their direct detection in vivo. Despite the plethora of data, those methods are still lacking. The autofluorescence from the extended beta-sheets of amyloids is here used to track fibrillation of S. cerevisiae Golgi Reassembly and Stacking Protein (Grh1). Grh1 has been implicated in starvation-triggered unconventional protein secretion (UPS), and here its participation also in heat shock response (HSR) is suggested. Fluorescence Lifetime Imaging (FLIM) is used to detect fibril autofluorescence in cells (E. coli and yeast) under stress (starvation and higher temperature). The formation of Grh1 large complexes under stress is further supported by size exclusion chromatography and ultracentrifugation. The data show for the first time in vivo detection of amyloids without the use of extrinsic probes as well as bring new perspectives on the participation of Grh1 in UPS and HSR. (AU)

• FAPESP's process: 15/50366-7 - Resolving mechanistic details of peptide transport across membranes using crystallographic and non-crystallographic structural biology approaches
• Grantee: Antonio José da Costa Filho
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/23863-2 - Molecular interactions of the Golgi Reassembly and Stacking Protein (GRASP) form Saccharomyces cerevisiae
• Grantee: Natália Aparecida Fontana
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 12/20367-3 - Structural and functional studies of the Golgi Re-Assembly and Stacking Protein (GRASP) from Cryptococcus neoformans
• Grantee: Antonio José da Costa Filho
• Support Opportunities: Regular Research Grants
• FAPESP's process: 18/24968-8 - Molecular interactions of the Saccharomyces cerevisiae golgin Bug1
• Grantee: Ariane Duarte Rosse
• Support Opportunities: Scholarships in Brazil - Scientific Initiation