title: Initial process of polyglutamine aggregate formation in vivo.
abstract: BACKGROUND
  Polyglutamine expansion in protein is responsible for several inherited neuro-degenerative diseases. The expansion has toxic effects on neural cells as well as results in forming aggregates. Using yeast, we examined the initial process of polyglutamine aggregate formation in vivo.
  
  
  RESULTS
  Following expression, polyglutamine tracts were of a soluble form during a lag period, and then formed insoluble complexes. The lag was prolonged and the formation of insoluble complex became slower by decreasing the number of polyglutamine tracts and by a treatment with guanidine hydrochloride. Gel filtration analysis revealed that the soluble polyglutamine existed in a small form. Formation of polyglutamine aggregates appeared to follow similar kinetics reported in the in vitro studies, where polyglutamine tracts self-aggregate in a length-, concentration- and time-dependent manner. However, in vivo, Hsp104 was required for the conversion from a soluble to an insoluble state. Without Hsp104, polyglutamine tracts tended to remain in a small soluble form, prolonging the lag. Moreover, the dependency on Hsp104 for aggregate formation was strong with the short polyglutamine tract, and decreased with the long polyglutamine tract.
  
  
  CONCLUSIONS
  For polyglutamine aggregate formation, a balance of parameters including the length of the polyglutamine tract, Hsp104, and level of polyglutamine expression determined its efficiency.
authors.affiliation: Department of Tumor Cell Biology, The Tokyo Metropolitan Institute of Medical Science, 3-18-22, Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan. ykimura@rinshoken.or.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Kimura
authors.forename: S
authors.initials: S
authors.lastname: Koitabashi
authors.forename: A
authors.initials: A
authors.lastname: Kakizuka
authors.forename: T
authors.initials: T
authors.lastname: Fujita
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D019791:Guanidine / Q000494:pharmacology; D006360:Heat-Shock Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D012995:Solubility
chemical_list: D005656:Fungal Proteins; D006360:Heat-Shock Proteins; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; C070972:HsP104 protein, S cerevisiae; C097188:polyglutamine; D019791:Guanidine