@article{mbs:/content/journal/micro/10.1099/mic.0.028399-0, author = "Uchida, Ikuo and Ishihara, Ryoko and Tanaka, Kiyoshi and Hata, Eiji and Makino, Sou-ichi and Kanno, Toru and Hatama, Shinichi and Kishima, Masato and Akiba, Masato and Watanabe, Atsushi and Kubota, Takayuki", title = "Salmonella enterica serotype Typhimurium DT104 ArtA-dependent modification of pertussis toxin-sensitive G proteins in the presence of [32P]NAD", journal= "Microbiology", year = "2009", volume = "155", number = "11", pages = "3710-3718", doi = "https://doi.org/10.1099/mic.0.028399-0", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.028399-0", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "PTX, pertussis toxin", keywords = "PNS, post nuclear supernatant", keywords = "MTC, mitomycin C", keywords = "LTX, heat-labile enterotoxin", keywords = "CHO, Chinese hamster ovary", keywords = "DT, definitive phage type", keywords = "CTX, cholera toxin", abstract = " Salmonella enterica serotype Typhimurium (S. Typhimurium) definitive phage type (DT) 104 has become a widespread cause of human and other animal infections worldwide. The severity of clinical illness in S. Typhimurium DT104 outbreaks suggests that this strain possesses enhanced virulence. ArtA and ArtB – encoded by a prophage in S. Typhimurium DT104 – are homologues of components of pertussis toxin (PTX), including its ADP-ribosyltransferase subunit. Here, we show that exposing DT104 to mitomycin C, a DNA-damaging agent, induced production of prophage-encoded ArtA/ArtB. Pertussis-sensitive G proteins were labelled in the presence of [32P]NAD and ArtA, and the label was released by HgCl2, which is known to cleave cysteine-ADP-ribose bonds. ADP-dependent modification of G proteins was markedly reduced in in vitro-synthesized ArtA6Arg-Ala and ArtA115Glu-Ala, in which alanine was substituted for the conserved arginine at position 6 (necessary for NAD binding) and the predicted catalytic glutamate at position 115, respectively. A cellular ADP-ribosylation assay and two-dimensional electrophoresis showed that ArtA- and PTX-induced ADP-ribosylation in Chinese hamster ovary (CHO) cells occur with the same type of G proteins. Furthermore, exposing CHO cells to the ArtA/ArtB-containing culture supernatant of DT104 resulted in a clustered growth pattern, as is observed in PTX-exposed CHO cells. Hydrogen peroxide, an oxidative stressor, also induced ArtA/ArtB production, suggesting that these agents induce in vivo synthesis of ArtA/ArtB. These results, taken together, suggest that ArtA/ArtB is an active toxin similar to PTX.", }