10.1371/journal.pgen.1005390.g001
Zhanna Lipatova
Zhanna
Lipatova
Nava Segev
Nava
Segev
The general Atgs Atg1 and Atg8 and the selective Atg Atg11, but not other selective Atgs, are required for macro-ER-phagy.
Public Library of Science
2015
erad
ER stress
degradation
ERQC
sequential steps
core Atgs
autophagy
use autophagic organelles
Ypt GTPases
ER membrane
upr
ap
system shuttles misfolded proteins
vacuole
role
pathway
ER quality control
2015-07-16 02:50:24
Figure
https://plos.figshare.com/articles/figure/_The_general_Atgs_Atg1_and_Atg8_and_the_selective_Atg_Atg11_but_not_other_selective_Atgs_are_required_for_macro_ER_phagy_/1485804
<p><b>A.</b> GFP-Snc1-PEM protein accumulates in <i>atg1∆</i>, <i>atg8∆</i>, and <i>atg11∆</i> mutant cells. Lysates were prepared from wild type (WT), <i>ypt1-1</i> (for comparison), <i>atg1∆</i>, <i>atg8∆</i>, and <i>atg11∆</i> mutant cells transformed with a 2μ plasmid expressing GFP-Snc1-PEM from the TPI promoter. The level of GFP-Snc1-PEM was determined using immunoblot analysis with anti-GFP antibodies. The bands were quantified and increase in the protein level in mutant versus the WT cells is shown under the blot and adjusted to the G6PDH loading control. <b>B.</b> GFP-Snc1-PEM protein accumulates in aberrant ER structures in <i>atg1∆</i>, <i>atg8∆</i>, and <i>atg11∆</i> mutant cells. The ER-marker Sec61 was tagged with mCherry in strains from panel A, and the cells were examined by live-cell microscopy. Shown from left to right: DIC, GFP, mCherry, merge, % cells with intracellular GFP-Snc1-PEM (number of cells with internal GFP-Snc1-PEM / number of cells visualized), and % cells in which intra-cellular Snc1-PEM co-localizes with Sec61. <b>C.</b> UPR is induced in <i>atg1∆</i>, <i>atg8∆</i>, and <i>atg11∆</i> mutant cells. Cells from panel A were transformed with a second plasmid that expresses β-gal from a UPR promoter. UPR was determined and expressed as % of the WT response. <b>D-E.</b> Unlike deletion of Atg11, deletion of other known selective Atgs required for the CVT pathway (Atg19), mitophagy (Atg32) and pexophagy (Atg36), does not result in increase of GFP-Snc1-PEM protein level (<b>D</b>), intra-cellular accumulation of GFP-Snc1-PEM, (<b>E</b>), and induction of the UPR response (<b>F</b>). Wild type (WT), <i>atg19∆</i>, <i>atg11∆</i>, <i>atg32∆</i>, and <i>atg36∆</i> mutant cells overexpressing GFP-Snc1-PEM were analyzed as described for panels A-C, respectively. <b>E.</b> Shown from left to right: DIC, GFP, and % cells with intracellular Snc1-PEM structures. +/- and error bars represent STDEV. Results in this figure represent at least two independent experiments.</p>