Selective amplification of an mRNA and related pseudogene for a human ADP-ribosylation factor, a guanine nucleotide-dependent protein activator of cholera toxin.

Abstract

ADP-ribosylation factors (ARFs) are =20-kDa proteins that act as GTP-dependent allosteric activators of cholera toxin. With deoxyinosine-containing degenerate oligonucleotide primers corresponding to conserved GTP-binding domains in ARFs, the polymerase chain reaction (PCR) was used to amplify simultaneously from human DNA portions of three ARF genes that include codons for 102 amino acids, with intervening sequences. Amplification products that differed in size because of differences in intron sizes were separated by agarose gel electrophoresis. One amplified DNA contained no introns and had a sequence different from those of known ARFs. Based on this sequence, selective oligonucleotide probes were prepared and used to isolate clone 'IARF 4, a putative

Differential interaction of ADP-ribosylation factors 1, 3, and 5 with rat brain Golgi membranes.

“…Based on immunoreactivity and assay of ARF activity, individual ARFs 1,3, and 5 appeared to behave independently and selectively in their GTP-dependent assoction with Golgi in vitro.…”

“…Mammalian ADP-ribosylation factors (ARFs), 20-kDa GTPbinding proteins, are products ofat least six ARF genes (1)(2)(3)(4)(5)(6). The recombinant proteins expressed in Escherichia coli exhibit the characteristic GTP-dependent stimulation ofcholera toxin A subunit (CTA)-catalyzed ADP-ribosylation (7).…”

“…Their deduced amino acid sequences contain consensus sequences that are believed to be involved in guanine nucleotide binding and GTP hydrolysis (8). Based on size, deduced amino acid sequence, and gene structure, they have been divided into three classes: class I, ARFs 1, 2, and 3; class II, ARFs 4 and 5; class III, ARF 6 (2,3,6,9). They are ubiquitous, highly conserved proteins distributed widely in eukaryotes from Giardia to mammals (10)(11)(12).…”

Selective amplification of additional members of the ADP-ribosylation factor (ARF) family: cloning of additional human and Drosophila ARF-like genes.

“…The first ARF, bovine ARF1, was cloned by screening cDNA libraries with oligonucleotide probes based on partial sequence information (9). A variety of other techniques have now been used to clone members of the ARF family, including functional screens (10)(11)(12), low-stringency hybridizations (9,13,14), PCR methodologies (15), and serendipity (16,17). At least 18 cDNA sequences ofmembers ofthe ARFfamily from at least nine different organisms have been reported.…”

Assignment<footref rid="foot01">¹</footref> of the human ADP-ribosylation factor-like 3 (ARL3) gene to chromosome 10 band q23.3 by radiation hybrid mapping

“…ADP-ribosylation factors (ARFs) are guanine nucleotidebinding proteins, approximately 20 kDa in size, that function as GTP-dependent allosteric activators of cholera toxin ADPribosyltransferase activity (Bobak et al, 1989, Monaco et al, 1990. They are thought to be involved in membrane trafficking and protein secretion (Hosaka et al, 1996).…”