Methods for the in vivo and in vitro analysis of [Het-s] prion infectivity.
Prions have been described in mammals and fungi. The [Het-s] infectious genetic element of the filamentous fungus Podospora anserina is the prion form of the HET-s protein. This protein is involved in the control of a cell death reaction termed heterokaryon incompatibility. The infectious form of HET-s corresponds to a self-perpetuating amyloid. The purpose of the present paper is to describe the techniques that can be used to analyse [Het-s] prion propagation in vivo and HET-s amyloid aggregation in vitro. In addition, we report several methods that can be used to infect Podospora with recombinant HET-s amyloid.
Benkemoun
Laura
L
Laboratoire de Génétique Moléculaire des Champignons, IBGC UMR CNRS 5095, Université de Bordeaux 2, Bordeaux, France.
Sabaté
Raimon
R
Malato
Laurent
L
Dos Reis
Suzana
S
Dalstra
Henk
H
Saupe
Sven J
SJ
Maddelein
Marie-Lise
ML
D000682:Amyloid / Q000737:chemistry / Q000648:ultrastructure; D002942:Circular Dichroism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008854:Microscopy, Electron; D048468:Podospora / Q000235:genetics / Q000378:metabolism*; D011328:Prions / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013172:Spores, Fungal / Q000235:genetics / Q000378:metabolism
D000682:Amyloid; D005656:Fungal Proteins; C083891:HET-S protein, Podospora anserina; D011328:Prions; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins