TABLE 3.
Intracellular localization of WT, A53T, and A30P α-Syn inSEC+,sec9, andsec14 cells
| α-Syn isoform |
Plasma membrane |
Plasma membrane aggregates |
Cytoplasm | Cytoplasmic aggregates |
|---|---|---|---|---|
| SEC+ | ||||
| GFP | ||||
| 23° | 0 | 0 | 80 | 0 |
| 37° | 13 | 0 | 174 | 0 |
| WT | ||||
| 23° | 36 | 0 | 33 (9) | 0 |
| 37° | 135 | 0 | 25 | 0 |
| A53T | ||||
| 23° | 22 | 0 | 8 | 0 |
| 37° | 72 | 0 | 9 (1) | 0 |
| A30P | ||||
| 23° | 1 | 0 | 99 (2) | 0 |
| 37° | 1 | 0 | 160 (1) | 0 |
| sec9 | ||||
| GFP | ||||
| 23° | 0 | 0 | 107 (2) | 0 |
| 37° | 0 | 0 | 96 (15) | 0 |
| WT | ||||
| 23° | 27 | 0 | 27 (7) | 1 |
| 37° | 18 | 16 | 31 (9) | 0 |
| A53T | ||||
| 23° | 71 | 5 | 47 (8) | 0 |
| 37° | 16 | 9 | 23 (10) | 0 |
| A30P | ||||
| 23° | 0 | 0 | 49 (6) | 0 |
| 37° | 0 | 0 | 40 (3) | 3 |
| sec14 | ||||
| GFP | ||||
| 23° | 0 | 0 | 36 | 0 |
| 37° | 0 | 0 | 30 (9) | 0 |
| WT | ||||
| 23° | 39 | 1 | 9 (2) | 0 |
| 37° | 9 | 8 | 10 | 3 |
| A53T | ||||
| 23° | 114 | 0 | 10 (1) | 0 |
| 37° | 10 | 15 | 15 | 14 |
| A30P | ||||
| 23° | 0 | 0 | 32 (9) | 1 |
| 37° | 0 | 0 | 39 (6) | 0 |
Yeast strains LRB906 (SEC+), LRB934 (sec9), and LRB933 (sec14), expressing GFP alone or GFP-α-Syn fusion proteins as indicated, were pregrown at 23° in 2% raffinose to early log phase. Galactose was then added at a final concentration of 2%, and cells were cultivated for an additional 2 hr at either 23° or 37°. Plasma membrane, uniform enhancement of fluorescence at cell periphery; plasma membrane aggregates, enhancement of peripheral fluorescence characterized by the presence of punctate foci; cytoplasm, fluorescence restricted to cytosol and internal organelles (those cells exhibiting enhanced nuclear fluorescence are indicated in parentheses); cytoplasmic aggregates, fluorescence restricted to the cytoplasm with one or more punctate foci.