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. 2018 Apr 30;109(1):41–62. doi: 10.1111/mmi.13966

Figure 6.

Figure 6

Lack of Apj1 expression, but not any of 11 other cytosolic J‐proteins, impairs Hsp104 curing of strong [PSI +]STR.

A. Strong [PSI +]STR bearing cells of the W303 genetic background lacking individual J‐proteins were passaged onto rich medium (left columns). Cells were then transformed with a plasmid overexpressing Hsp104 (GPD‐HSP104) that normally cures [PSI +] (right columns). Color phenotype assays are shown for representative transformants: for apj1‐Δ 48 out of 90 transformants remained [PSI +]; for all other strains curing was complete with n ≥ 10.

B. Lysates of a wild‐type strain (wt), a strain lacking Apj1 expression (apj1‐Δ), a strain lacking Caj1 expression (caj1‐Δ) and a strain lacking the J‐domain of Cwc23 (cwc23‐ΔJ) were resolved by SDS‐PAGE and subjected to immunoblot analysis using antibodies specific for Hsp104, Ssa1–4 or Sis1.

C. [PSI +]STR cells with a deletion of the APJ1 gene (apj1‐Δ) were transformed first by plasmids overexpressing one of the two J‐proteins (↑Apj1 or ↑Sis1) or empty vector (vector), followed by a subsequent transformation with plasmid overexpressing Hsp104 (GPD‐HSP104) that normally cures [PSI +]. Color phenotype assays are shown for representative transformants (n ≥ 10 for each variant).

D. sis1‐Δ cells bearing [PSI +]STR and expressing Sis1‐ΔG/F from a plasmid were transformed with either empty vector (top row) or plasmid overexpressing Apj1 (bottom row) and subsequently transformed with GPD‐HSP104.