Molecular Characterization of a Conserved, Guanine Nucleotide-Dependent ADP-Ribosylation Factor in Drosophila melanogaster

James J. Murtagh, Fang Jen S. Lee, Peter Deak, Linda M. Hall, Lucia Monaco, Chii Ming Lee, Linda A. Stevens, Joel Moss, Martha Vaughan

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Abstract

ADP-Ribosylation factors (ARFs) are ubiquitous ∼ 20-kDa guanine nucleotide-binding proteins that stimulate cholera toxin-catalyzed ADP-ribosylation in vitro. Because the functional role(s) of ARF in mammalian systems is (are) elusive, we looked for ARF in Drosophila melanogaster, and report the partial purification and molecular cloning of an ARF from Drosophila. We cloned the Drosophila ARF 1 gene without library screening by a combination of 5 polymerase chain reactions (PCRs), yielding a 546-base open reading frame encoding 182 amino acids, which are >93% identical to those of mammalian class I ARFs. This ARF gene maps to 79F3-6 in the proximal region of the left arm of Drosophila chromosome 3. The Drosophila ARF 1 gene structure, including placement of introns, is highly conserved relative to mammalian class 1 ARF genes. A single ARF mRNA species of 1.8 kb was abundant in all Drosophila body segments. Recombinant Drosphila ARF 1 synthesized in Escherichia coli had biochemical and immunochemical activities similar to those of mammalian ARF. The similarities of sequence and biochemical properties between Drosophila and mammalian ARFs contrast with their differences from Drosophila arl (ARF-like protein), which does not stimulate cholera toxin-catalyzed ADP-ribosylation, and is only ∼52–56% identical in amino acid sequence to mammalian ARFs.

Original languageEnglish
Pages (from-to)6011-6018
Number of pages8
JournalBiochemistry
Volume32
Issue number23
DOIs
Publication statusPublished - Jan 1 1993

ASJC Scopus subject areas

  • Biochemistry

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    Murtagh, J. J., Lee, F. J. S., Deak, P., Hall, L. M., Monaco, L., Lee, C. M., Stevens, L. A., Moss, J., & Vaughan, M. (1993). Molecular Characterization of a Conserved, Guanine Nucleotide-Dependent ADP-Ribosylation Factor in Drosophila melanogaster. Biochemistry, 32(23), 6011-6018. https://doi.org/10.1021/bi00074a012