Molecular cloning and disruption of a novel gene encoding UDP-glucose: tetrahydrobiopterin alpha-glucosyltransferase in the cyanobacterium Synechococcus sp. PCC 7942

FEBS Lett. 2001 Aug 3;502(3):73-8. doi: 10.1016/s0014-5793(01)02667-9.

Abstract

The gene encoding UDP-glucose:tetrahydrobiopterin alpha-glucosyltransferase (BGluT) was cloned from the genomic DNA of Synechococcus sp. PCC 7942. The encoded protein consisting of 359 amino acid residues was verified in vitro and in vivo to be responsible for the synthesis of tetrahydrobiopterin (BH4)-glucoside produced in the organism. The BGluT gene is the first cloned in pteridine glycosyltransferases and also a novel one cloned so far in UDP-glycosyltransferases. The mutant cells disrupted in the BGluT gene produced only aglycosidic BH4 at a level of 8.3% of the BH4-glucoside in wild type cells and exhibited half of the wild type growth in normal photoautotrophic conditions. These results suggest that the glucosylation of BH4 is required for the maintenance of the high cellular concentration of the compound, thereby supporting the normal growth of Synechococcus sp. PCC 7942.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aclarubicin
  • Amino Acid Sequence
  • Antineoplastic Combined Chemotherapy Protocols
  • Bacterial Proteins*
  • Base Sequence
  • Biopterins / analogs & derivatives*
  • Biopterins / biosynthesis
  • Biopterins / metabolism*
  • Blotting, Southern
  • Blotting, Western
  • Cell Division
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Cyanobacteria / enzymology*
  • Cytarabine
  • DNA Primers / chemistry
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Genes, Bacterial
  • Glucosyltransferases / chemistry
  • Glucosyltransferases / genetics*
  • Glucosyltransferases / metabolism
  • Glycosylation
  • Molecular Sequence Data
  • Mutation
  • Polymerase Chain Reaction
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • DNA Primers
  • DNA, Bacterial
  • Recombinant Proteins
  • Cytarabine
  • Biopterins
  • Aclarubicin
  • BGluT protein, Synechococcus sp. PCC 7942
  • Glucosyltransferases
  • sapropterin

Supplementary concepts

  • BGM 84 protocol

Associated data

  • GENBANK/AJ003064