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.