In:
European Journal of Biochemistry, Wiley, Vol. 259, No. 1-2 ( 1999-01), p. 112-119
Kurzfassung:
The GTS1 gene product, Gts1p, has pleiotropic effects on the timing of budding, cell size, heat tolerance, sporulation and the lifespan of the yeast Saccharomyces cerevisiae . In this study, we found (using the yeast two‐hybrid system) that Gts1p forms homodimers throughout the 18‐amino acid region 296–313 which has considerable similarity to a region downstream of the Walker nucleotide‐binding motif A of some ATP‐binding cassette (ABC) transporters. The region contains two aspartic acid residues at 301 and 310 preceded by hydrophobic amino acid residues, and Gts1p with an Asp310 to Ala substitution showed considerably reduced homodimerization, as shown by the two‐hybrid assay. Overexpression of the point‐mutated Gts1p did not efficiently induce the Gts1p‐related phenotypes described above, suggesting that the homodimerization of Gts1p is required for it to function in vivo . The C‐terminal cytoplasmic domain of the yeast ABC transporters Mdl1p (multidrug resistance‐like transporter) and Ycf1p (yeast cadmium factor or glutathione S‐conjugate pump) bound to Gts1p in the two‐hybrid system, and the heterodimerization activity of the Gts1p with the Asp301 to Ala substitution was more affected than the Gts1p with the Asp310 to Ala substitution. Overexpression of GTS1 considerably reduced, and disruption of GTS1 slightly decreased, cellular resistance to cycloheximide, cadmium, cisplatin and 1‐chloro‐2,4‐dinitrophenol, which (except for cycloheximide) are all substrates of Ycf1p. These results suggest that Gts1p interacts with some ABC transporters through the binding site overlapping that of homodimerization and modulates their activity.
Materialart:
Online-Ressource
ISSN:
0014-2956
,
1432-1033
DOI:
10.1046/j.1432-1327.1999.00008.x
Sprache:
Englisch
Verlag:
Wiley
Publikationsdatum:
1999
ZDB Id:
1398347-7
ZDB Id:
2172518-4
SSG:
12
