Deletion of the cnxE gene encoding the gephyrin-like protein involved in the final stages of molybdenum cofactor biosynthesis in Aspergillus nidulans.
The Aspergillus nidulans cnxE gene, required for molybdenum cofactor biosynthesis, was isolated by functional complementation of an Escherichia coli mogA mutant strain. The deduced CnxE polypeptide consists of two domains which display similarity to the E. coli proteins MoeA and MogA, respectively, separated by a putative hinge region of around 58 amino acid residues which is notably histidine rich. A deletion mutant lacking the entire cnxE gene, including both MoeA-like and MogA-like domains, was identified. Compared to the wild type, a small increase in the intermediate precursor Z was observed in the deletion strain but was significant only under conditions in which the molybdoenzyme nitrate reductase was induced. Elevated levels of the pathway intermediate molybdopterin were found both under nitrate reductase-inducing and non-inducing conditions in the deletion mutant compared to the wild type. This increase is in contrast to previous results for cnxABC, cnxF, cnxG, and cnxH mutants, in which the levels of molybdopterin were substantially reduced, and therefore supports previously published classical genetic and biochemical studies that indicated that the CnxE protein is likely to be involved in the final stages of molybdenum cofactor biosynthesis. We have found no evidence during our chemical analysis for any involvement of this protein in the intermediate section of the molybdenum cofactor biosynthetic pathway (i.e. in the synthesis of molybdopterin from precursor Z), as has been suggested previously for E. coli MoeA. The 2.5-kb cnxE transcript is not abundant and appears to be expressed constitutively.
Millar
L J
LJ
School of Biology, University of St. Andrews, St. Andrews, Fife KY16 9TH, UK.
Heck
I S
IS
Sloan
J
J
Kana'n
G J
GJ
Kinghorn
J R
JR
Unkles
S E
SE
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D015139:Blotting, Southern; D002352:Carrier Proteins; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D003067:Coenzymes; D018076:DNA, Complementary; D004271:DNA, Fungal / Q000737:chemistry; D004792:Enzyme Precursors / Q000096:biosynthesis*; D017353:Gene Deletion; D005786:Gene Expression Regulation; D008565:Membrane Proteins; D008667:Metalloproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D008982:Molybdenum / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D009097:Multienzyme Complexes / Q000235:genetics*; D009154:Mutation; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000378:metabolism; D010957:Plasmids; D011621:Pteridines / Q000378:metabolism*
D002352:Carrier Proteins; D003067:Coenzymes; D018076:DNA, Complementary; D004271:DNA, Fungal; D004792:Enzyme Precursors; D008565:Membrane Proteins; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D009097:Multienzyme Complexes; D050901:Nitrate Reductase; D009565:Nitrate Reductases; D011621:Pteridines; C073441:gephyrin; C022261:molybdopterin