Characterization of an ADP-ribosylation factor-like 1 protein in Saccharomyces cerevisiae
Journal
Journal of Biological Chemistry
Journal Volume
272
Journal Issue
49
Pages
30998-31005
Date Issued
1997
Author(s)
Abstract
ADP-ribosylation factors (ARFs) are highly conserved ~20-kDa guanine nucleotide-binding proteins that enhance the ADP-ribosyltransferase activity of cholera toxin and are believed to participate in vesicular transport in both exocytic and endocytic pathways. Several ARF-like proteins (ARLs) have been cloned from Drosophila, rat, and human; however, the biological functions of ARLs are unknown. We have identified a yeast gene (ARL1) encoding a protein that is structurally related (>60% identical) to human, rat, and Drosophila ARLL Biochemical analyses of purified recombinant yeast ARL1 (yARL1) protein revealed properties similar in those ARF and ARL1 proteins, including the ability to bind and hydrolyze GTP. Like other ARLs, recombinant yARL1 protein did not stimulate cholera toxin-catalyzed auto- ADP-ribosylation. yARL1 was not recognized by antibodies against mammalian ARLs or yeast ARFs. Anti-yARL1 antibodies did not cross-react with yeast ARFs, but did react with human ARLs. On subcellular fractionation, yARL1, similar to yARF1, was localized to the soluble fraction. The amine terminus of yARL1, like that of ARF, was myristoylated. Unlike Drosophila Arl1, yeast ARL1 was not essential for cell viability. Like rat ARL1, yARL1 might be associated in part with the Golgi complex. However, yARL1 was not required for endoplasmic reticulum-to-Golgi protein transport, and it may offer an opportunity to define an ARL function in another kind of vesicular trafficking, such as the regulated secretory pathway.
SDGs
Other Subjects
amino terminal sequence; animal cell; antigen antibody complex; article; drosophila melanogaster; endocytosis; exocytosis; gene expression; myristylation; nonhuman; polymerase chain reaction; priority journal; protein analysis; protein purification; saccharomyces cerevisiae; yeast; ADP-Ribosylation Factors; Amino Acid Sequence; Animals; Biological Transport; Carrier Proteins; Endoplasmic Reticulum; Fluorescent Antibody Technique, Indirect; Golgi Apparatus; GTP Phosphohydrolases; Guanosine 5'-O-(3-Thiotriphosphate); Humans; Membrane Proteins; Molecular Sequence Data; Myristic Acid; Rats; Recombinant Proteins; Saccharomyces cerevisiae; Sequence Alignment; Animalia; Drosophila melanogaster; Mammalia; Melanogaster; Saccharomyces cerevisiae
Type
journal article