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J. Biol. Chem., Vol. 266, Issue 5, 2772-2777, 02, 1991
M Tsuchiya, SR Price, SC Tsai, J Moss and M Vaughan
ADP-ribosylation factors (ARFs) are approximately 20-kDa guanine
nucleotide-binding proteins that serve as GTP-dependent allosteric
activators of cholera toxin ADP-ribosyltransferase activity. Four species
of mammalian ARF, termed ARF 1-4, have been identified by cloning.
Hybridization of a bovine ARF 2 cDNA under low stringency with mammalian
poly(A)+ RNA resulted in multiple bands that were subsequently assigned to
the known ARF genes using ARF-specific oligonucleotide probes. The relative
signal intensities of some bands (e.g. the 3.8- and 1.3-kilobase (kb)
mRNAs) that hybridized with the cDNA were not, however, consistent with the
intensities observed with the individual ARF-specific oligonucleotide
probes. These inconsistencies suggested that other ARF-like mRNAs were
comigrating with known ARF mRNAs. To explore this possibility, a cyclic
AMP- differentiated HL-60 Lambda ZAP library was screened using the bovine
ARF 2 cDNA. Clones corresponding to known ARF genes (1, 3, and 4) were
identified by hybridization of positive clones with oligonucleotide probes
specific for each ARF species; ARF 2 cDNA-positive,
oligonucleotide-negative clones were sequenced. Two new ARF-like genes, ARF
5 and 6, encoding proteins of 180 and 175 amino acids, respectively, were
identified. Both proteins contain consensus sequences believed to be
involved in guanine nucleotide binding and GTP hydrolysis. ARF 5 was most
similar in deduced amino acid sequence to ARF 4, which also has 180 amino
acids. ARF 6, whose deduced amino acid sequence is identical with that of a
putative chicken pseudogene (CPS1) except for a serine/threonine
substitution, was different from other ARF species in size and deduced
amino acid sequence. With mammalian poly(A)+ RNA from a variety of tissues
and cultured cells, ARF 5 preferentially hybridized with a 1.3-kb mRNA,
whereas ARF 6 hybridized with 1.8- and 4.2-kb mRNAs. The fact that the
sizes of these mRNAs are similar to those of other ARFs (ARF 1, 1.9 kb; ARF
2, 2.6 kb; ARF 3, approximately 3.8 and 1.3 kb; ARF 4, 1.8 kb) explain the
previously observed inconsistencies between the cDNA and ARF-specific
oligonucleotide hybridization patterns. All six ARF cDNAs are more similar
to each other than to other approximately 20-kDa guanine nucleotide-binding
proteins.
Molecular identification of ADP-ribosylation factor mRNAs and their expression in mammalian cells
Laboratory of Cellular Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
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R. R. Dana, C. Eigsti, K. L. Holmes, and T. L. Leto A Regulatory Role for ADP-ribosylation Factor 6 (ARF6) in Activation of the Phagocyte NADPH Oxidase J. Biol. Chem., October 13, 2000; 275(42): 32566 - 32571. [Abstract] [Full Text] [PDF]
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N. Vitale, G. Pacheco-Rodriguez, V. J. Ferrans, W. Riemenschneider, J. Moss, and M. Vaughan Specific Functional Interaction of Human Cytohesin-1 and ADP-ribosylation Factor Domain Protein (ARD1) J. Biol. Chem., July 7, 2000; 275(28): 21331 - 21339. [Abstract] [Full Text] [PDF]
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