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. 2004 Apr 15;379(Pt 2):243–251. doi: 10.1042/BJ20031585

Inosine 5'-monophosphate dehydrogenase binds nucleic acids in vitro and in vivo.

Jeremy E McLean 1, Nobuko Hamaguchi 1, Peter Belenky 1, Sarah E Mortimer 1, Martin Stanton 1, Lizbeth Hedstrom 1
PMCID: PMC1224093  PMID: 14766016

Abstract

Inosine 5'-monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in the de novo biosynthesis of guanine nucleotides. In addition to the catalytic domain, IMPDH contains a subdomain of unknown function composed of two cystathione beta-synthase domains. Our results, using three different assays, show that IMPDHs from Tritrichomonas foetus, Escherichia coli, and both human isoforms bind single-stranded nucleic acids with nanomolar affinity via the subdomain. Approx. 100 nucleotides are bound per IMPDH tetramer. Deletion of the subdomain decreases affinity 10-fold and decreases site size to 60 nucleotides, whereas substitution of conserved Arg/Lys residues in the subdomain with Glu decreases affinity by 20-fold. IMPDH is found in the nucleus of human cells, as might be expected for a nucleic-acid-binding protein. Lastly, immunoprecipitation experiments show that IMPDH binds both RNA and DNA in vivo. These experiments indicate that IMPDH has a previously unappreciated role in replication, transcription or translation that is mediated by the subdomain.

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