Impact of Mycoplasma hyorhinis infection on L-arginine metabolism: differential regulation of the human and murine iNOS gene.
Infection with mycoplasma is a common problem in cell cultures, with Mycoplasma hyorhinis being the predominant species. Here we investigate the effect of M. hyorhinis infection on L-arginine metabolism, with focus on iNOS-mediated NO synthesis in murine keratinocytes and the human colon cancer cell line DLD-1. iNOS and arginase are L-arginine-metabolizing enzymes involved in the regulation of inflammatory processes, with NO contributing to innate immunity. In murine cells, M. hyorhinis infection enhances cytokine-induced iNOS expression and augments iNOS activity, whereas in the absence of cytokines it causes de novo induction of iNOS mRNA without subsequent translation into iNOS protein. In turn, arginase-1 mRNA expression is diminished in M. hyorhinis-infected murine keratinocytes, resulting in decreased arginase activity. One of the underlying upstream mechanisms is NF-kappaB activation. In contrast, in human cells neither iNOS mRNA nor protein expression is affected by M. hyorhinis infection, but NO synthesis is enhanced, which may be caused by increased L-arginine import. This demonstrates that infection with M. hyorhinis leads to different effects on gene regulation of the murine and human iNOS gene. Our study underlines the importance of routine checking of cell cultures for mycoplasma contamination, particularly in studies on NO-mediated effects or inflammatory processes.
Kagemann
Guido
G
Institute for Biochemistry and Molecular Biology I, Heinrich-Heine-University of Düsseldorf, Universitätsstr. 1, D-40225 Düsseldorf, Germany.
Henrich
Birgit
B
Kuhn
Michaela
M
Kleinert
Hartmut
H
Schnorr
Oliver
O
D000818:Animals; D001119:Arginase / Q000235:genetics; D001120:Arginine / Q000378:metabolism*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D015603:Keratinocytes / Q000201:enzymology / Q000382:microbiology; D051379:Mice; D045790:Mycoplasma hyorhinis / Q000302:isolation & purification / Q000502:physiology*; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism
D016328:NF-kappa B; D012333:RNA, Messenger; D004247:DNA; D001120:Arginine; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D001119:Arginase