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J. Biol. Chem., Vol. 277, Issue 35, 31373-31380, August 30, 2002
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§,
From the In Escherichia coli there are two
pathways for conversion of adenine into guanine nucleotides, both
involving the intermediary formation of IMP. The major pathway involves
conversion of adenine into hypoxanthine in three steps via adenosine
and inosine, with subsequent phosphoribosylation of hypoxanthine to
IMP. The minor pathway involves formation of ATP, which is converted
via the histidine pathway to the purine intermediate
5-amino-4-imidazolecarboxamide ribonucleotide and, subsequently, to
IMP. Here we describe E. coli mutants, in which a third
pathway for conversion of adenine to IMP has been activated. This
pathway was shown to involve direct deamination of adenine to
hypoxanthine by a manganese-dependent adenine deaminase
encoded by a cryptic gene, yicP, which we propose be
renamed ade. Insertion elements, located from
Department of Biological Chemistry,
Institute of Molecular Biology, University of Copenhagen, Sølvgade
83H, DK1307 Copenhagen K, ¶ John. F. Kennedy Institute, Gl.
Landevej 7, 2600 Glostrup,
Department of Biochemistry and
Molecular Biology, University of Southern Denmark, Campusvej 55,
DK-5230 Odense M, Denmark
145 to +13
bp relative to the transcription start site, activated the
ade gene as did unlinked mutations in the hns
gene, encoding the histone-like protein H-NS. Gene fusion analysis
indicated that ade transcription is repressed more than
10-fold by H-NS and that a region of 231 bp including the
ade promoter is sufficient for this regulation. The
activating insertion elements essentially eliminated the
H-NS-mediated silencing, and stimulated ade gene expression
2-3-fold independently of the H-NS protein.
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