ADP-ribosyltransferases including toxins secreted by Vibrio cholera, Pseudomonas aerurginosa, and other pathogenic bacteria inactivate the function of human target proteins by attaching ADP-ribose onto a critical amino acid residue. Cross-species polymerase chain reaction (PCR) and database mining identified the orthologs of these ADP-ribosylating toxins in humans and the mouse. The human genome contains four functional toxin-related ADP-ribosyltransferase genes (ARTs) and two related intron-containing pseudogenes; the mouse has six functional orthologs. The human and mouse ART genes map to chromosomal regions with conserved linkage synteny. The individual ART genes reveal highly restricted expression patterns, which are largely conserved in humans and the mouse. We confirmed the predicted extracellular location of the ART proteins by expressing recombinant ARTs in insect cells. Two human and four mouse ARTs contain the active site motif (R-S-EXE) typical of arginine-specific ADP-ribosyltransferases and exhibit the predicted enzyme activities. Two other human ARTs and their murine orthologues deviate in the active site motif and lack detectable enzyme activity. Conceivably, these ARTs may have acquired a new specificity or function. The position-sensitive iterative database search program PSI-BLAST connected the mammalian ARTs with most known bacterial ADP-ribosylating toxins. In contrast, no related open reading frames occur in the four completed genomes of lower eucaryotes (yeast, worm, fly, and mustard weed). Interestingly, these organisms also lack genes for ADP-ribosylhydrolases, the enzymes that reverse protein ADP-ribosylation. This suggests that the two enzyme families that catalyze reversible mono-ADP-ribosylation either were lost from the genomes of these nonchordata eucaryotes or were subject to horizontal gene transfer between kingdoms.Keywords: ADP-ribosylation; recombinant proteins; PSI-BLAST; orthologous genes; paralogous gene; cross-species PCR; database searches The purpose of this study was to identify all recognizable human and mouse members of the family of toxin-related mono-ADP-ribosyltransferases; to clone, sequence, and chromosomally map their genes and cDNAs; to express the gene products as recombinant proteins and assay their enzyme activities; and to assess structural and sequence similarities between bacterial and mammalian ADP-ribosyltransferases.ADP-ribosylation is an enzyme-catalyzed post-translational protein modification in which the ADP-ribose moiety is transferred from NAD+ to a specific amino acid in a Reprint requests to: Friedrich Koch-Nolte, Institute for Immunology, University Hospital, Martinistr. 52, D-20246 Hamburg, Germany, e-mail: nolte@uke.uni-hamburg.de; fax: 49-40-42803-4243.Abbreviations: ARH, ADP-ribosylhydrolase; GAPD, glyceraldehyde-3-phosphate dehydrogenase; GPI, glycosylphosphatidylinositol; HPRT, hypoxanthine phosphoribosyltransferase; mART, mono(ADP-ribosyl)transferase; NAD+, nicotine adenine dinucleotide; PBS, phosphate-buffered saline; pART, poly...
Two novel human members of an emerging mammalian gene family related to monoadp-ribosylating bacterial toxins Koch-Nolte, F.; Haag, F.; Braren, R.; Kuhl, M.; Hoovers, J.M.N.; Balasubramanian, S.; Bazan, F.; Thiele, H. Published in: Genomics DOI:10. 1006/geno.1996.4520 Link to publication Citation for published version (APA):Koch-Nolte, F., Haag, F., Braren, R., Kuhl, M., Hoovers, J. M. N., Balasubramanian, S., ... Thiele, H. (1997). Two novel human members of an emerging mammalian gene family related to mono-adp-ribosylating bacterial toxins. Genomics, 39, 370-376. DOI: 10.1006/geno.1996 General rights It is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. ribose moiety from NAD / to a specific amino acid in a Mono-ADP-ribosylation is one of the posttranslational target protein while the nicotinamide moiety is reprotein modifications regulating cellular metabolism, leased (Jacobson and Jacobson, 1989). Mono-ADP-riboe.g., nitrogen fixation, in prokaryotes. Several bacterial sylation was originally discovered as the mechanism toxins mono-ADP-ribosylate and inactivate specific pro-by which diphtheria toxin blocks protein synthesis teins in their animal hosts. Recently, two mammalian (Honjo et al., 1968). A number of other potent bacterial GPI-anchored cell surface enzymes with similar activi-toxins, including cholera, pertussis, and Escherichia ties were cloned (designated ART1 and ART2). We have coli heat labile enterotoxins, interfere with signal now identified six related expressed sequence tags transduction in human host cells by ADP-ribosylating (ESTs) in the public database and cloned the two novel regulatory G-proteins (Aktories, 1991; Moss and human genes from which these are derived (designated Vaughan, 1990). Since cellular protein functions are The suspicion that mono-ADP-ribosylation might genes to human chromosomes 4p14-p15.1 and 12q13.2-also be used as a mechanism to regulate endogenous q13.3. Northern blot analyses show that these genes are protein functions was first corroborated in photosynspecifically expressed in testis and spleen, respectively. thetic bacteria. Fixation of nitrogen is essential for all Comparison of genomic and cDNA sequences reveals a living organisms. In Rhodospirillium rubrum and conserved exon/intron structure, with an unusually large Azospiri...
Rt6 is a T cell-restricted GPI-anchored membrane protein and a member of the family of mono(ADP-ribosyl)transferases. One of the two murine Rt6 genes is deleted in NZW mice. This finding is reminiscent of the deletion of one of the TCR beta genes in the same mouse strain and it is an intriguing possibility that these gene deletions arose by a common genetic mechanism. The Rt6 locus retained by the NZW mouse (designated Rt6-1) is polymorphic among inbred strains of laboratory mice. The NZW mouse shows several strain-specific restriction fragment length variants in this Rt6 locus and five amino acid substitutions occur in the predicted native Rt6 polypeptide of the NZW mouse relative to the corresponding polypeptides of NZB and BALB/c mice. Whereas transcript levels of the two Rt6 genes appear to be normal in spleen and intestine of NZB mice, the corresponding tissues of NZW mice show reduced levels of transcripts from the Rt6 locus retained in this mouse strain. Moreover, reduced levels of Rt6 mRNA also occur in spleen and intestine of (NZB x NZW)F1 hybrid animals, indicating that F1 animals have inherited a dominant factor from the genetic background of the NZW mouse, resulting in low levels of Rt6 expression. It is conceivable that the alterations in the Rt6 genes of the NZW mouse and/or the factor(s) affecting defective Rt6 expression constitute part of the genetic contribution of the NZW mouse to the autoimmune lupus-like disease in (NZB x NZW)F1 animals.(ABSTRACT TRUNCATED AT 250 WORDS)
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