Abstract
The xpt and pbuX genes from Bacillus subtilis were cloned, and their nucleotide sequences were determined. The xpt gene encodes a specific xanthine phosphoribosyltransferase, and the pbuX gene encodes a xanthine-specific purine permease. The genes have overlapping coding regions, and Northern (RNA) blot analysis indicated an operon organization. The translation of the second gene, pbuX, was strongly dependent on the translation of the first gene, xpt. Expression of the operon was repressed by purines, and the effector molecules appear to be hypoxanthine and guanine. When hypoxanthine and guanine were added together, a 160-fold repression was observed. The regulation of expression was at the level of transcription, and we propose that a transcription termination-antitermination control mechanism similar to the one suggested for the regulation of the purine biosynthesis operon exists. The expression of the xpt-pbuX operon was reduced when hypoxanthine served as the sole nitrogen source. Under these conditions, the level of the hypoxanthine- and xanthine-degrading enzyme, xanthine dehydrogenase, was induced more than 80-fold. The xanthine dehydrogenase level was completely derepressed in a glnA (glutamine synthetase) genetic background. Although the regulation of the expression of the xpt-pbuX operon was found to be affected by the nitrogen source, it was normal in a glnA mutant strain. This result suggests the existence of different signalling pathways for repression of the transcription of the xpt-pbuX operon and the induction of xanthine dehydrogenase.
MeSH Terms
Amino Acid Sequence
Bacillus subtilis/enzymology,genetics
Bacterial Proteins
Base Sequence
Cloning, Molecular
Gene Expression Regulation, Bacterial/drug effects,physiology
Genes, Bacterial
Glutamate-Ammonia Ligase/metabolism
Membrane Transport Proteins/genetics
Molecular Sequence Data
Nitrogen/pharmacology
Nucleic Acid Conformation
Operon/genetics
Pentosyltransferases/genetics,metabolism
Protein Biosynthesis/genetics
Purines/metabolism,pharmacology
RNA, Bacterial/analysis,chemistry
RNA, Messenger/analysis,chemistry
Recombinant Fusion Proteins
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Transcription, Genetic/genetics
Xanthine
Xanthines/metabolism
Chemicals
Bacterial Proteins
Membrane Transport Proteins
PbuX protein, Bacillus subtilis
Purines
RNA, Bacterial
RNA, Messenger
Recombinant Fusion Proteins
Xanthines
Xanthine
Pentosyltransferases
xanthine phosphoribosyltransferase
glutamine synthetase I
Glutamate-Ammonia Ligase
Nitrogen
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Christiansen L C
Department of Biological Chemistry, University of Copenhagen, Denmark.
Schou S
Nygaard P
Saxild H H
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