Functional characterization and genetic study of an Arf-like protein, Arl3p in Saccharomyces cerevisiae
Date Issued
2004
Date
2004
Author(s)
Chang, Yi-Pin
DOI
en-US
Abstract
ADP-ribosylation factors (Arfs) are highly conserved small GTP-binding proteins involving vesicular membrane transport. To investigate biological function of a yeast Arf-like protein, Arl3p, yeast two-hybrid screens and synthetic lethal screens of ARL3 were performed. We found a novel protein, named Ali1p (Arf-like 3 interacting protein) that interacts with Arl3p in two-hybrid screen. The in vivo interaction of Ali1p and Arl3p was confirmed by GST-binding assay. Subcellular localization of Arl3p or Ali1p was examined. Arl3p co-localized with late Golgi marker, Sft2p. The Ali1p also showed Golgi-like or endosome-like punctate distribution. Arl3p seems not required for the punctate pattern of Ali1p, and vice versa. Interestingly, Arl3p lost its co-localization with Sft2p when Ali1p was over-expressed. arl3 and ali1 mutants both displayed Congo red sensitivity indicative of cell wall defect. However, arl3, but not ali1 mutant cells exhibited the abnormal distribution of a Gas1p which is involved in cell wall biogenesis, suggesting that Ali1p, different from Arl3p, may participate in distinct pathway with Arl3p to affect the cell wall biogenesis. Synthetic lethal screen of ARL3 was also performed and many candidate genes involved in different vesicular transport pathway were identified. By the colony- sector assay we showed that N-terminal 8 a. a. of Arl3p was essential for its function. Moreover, disruption of both ARL3 and COG8 result in the abnormal accumulation of a vital fluorescence dye, FM4-64, suggesting that Arl3p and Cog8p may be involved in vesicles targeting/fusion or vacuole biogenesis. This study may highlight many possible roles of Arl3p in different trafficking pathways.
Subjects
第三腺核苷
二磷酸醣化因子相似蛋白
ARL3 vesicle trafficking Arf
Type
other
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