A bipolar functionality of Q/N-rich proteins: Lsm4 amyloid causes clearance of yeast prions
Version of Record online: 20 MAR 2013
© 2013 The Authors. Microbiology Open published by John Wiley & Sons Ltd.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Volume 2, Issue 3, pages 415–430, June 2013
How to Cite
MicrobiologyOpen 2013; 2(3): 415–430
- Issue online: 7 JUN 2013
- Version of Record online: 20 MAR 2013
- Manuscript Accepted: 19 FEB 2013
- Manuscript Revised: 13 FEB 2013
- Manuscript Received: 2 JAN 2013
- The Ministry of Education, Sports, Culture, Science and Technology of Japan
Figure S1. Absence of the growth inhibition effect in [PSI+] or [URE3] cells harboring an LSM4-overexpressing plasmid on SC-Leu selective media. (A) Absence of the growth inhibition effect in [PSI+] cells. [PSI+][rnq−] cells (NPK50) were transformed with pRS425 (an empty vector) or pRS425GPDp-LSM4 (a multicopy LEU2+ plasmid expressing LSM4 under the control of the GPD promoter; denoted by pLSM4). Transformants were subsequently incubated on SC-Leu media for 3 days. (B) Absence of the growth inhibition effect in [URE3] cells. [URE3][rnq−] cells (NPK302) were subjected to the same experiment as (A).
Figure S2. Cellular abundance and thermotolerance activity of Hsp104 are unaffected by LSM4 expression. (A) Cellular abundance of Hsp104 on LSM4 overexpression. LSM4 was overexpressed from pRS425GPDp-LSM4 (under the GPD promoter) in NPK302 [URE3][rnq−] strain. Due to the low amount of lysate loaded, the Lsm4 protein expressed from the genome (see the lane of “Empty vector”) was narrowly observed. Immunoblotting was carried out using anti-Hsp104 antibody, anti-Lsm4 antibody, and anti-Pgk1 antibody. (B) Thermotolerance of LSM4-overexpressing cells. Cultures in the mid-log phase were incubated at 37°C for 1 h, and then heat treated at 50°C for 20 min. Survival rates were visualized by fivefold serial dilutions on YPD plate with untreated controls. Used strains were NPK301 ([ure-o][rnq−]) carrying an empty vector (top) or pRS425GPDp-LSM4 (middle), and NPK377 (NPK301-derivative carrying Δhsp104; bottom).
Table S1. PCR primers.
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