Sequence analysis, expression, and conservation of Escherichia coli uracil DNA glycosylase and its gene (ung).
The complete nucleotide sequence of the Escherichia coli ung gene is described. Transcription initiation and termination sites were determined by S1 nuclease and RNase mapping. The common prokaryotic -35, -10, and the ribosome binding site sequences are represented by TGTTCTGTA, TAAGCTA, and AGGAGAG at their respective locations. A putative hairpin transcription terminator structure is present at the major transcription terminator sites. The open reading frame of the ung gene codes for a protein of 229 amino acids (25,664 daltons). The molecular weight, amino acid composition, and the N-terminal amino acid sequence of the uracil DNA glycosylase purified from E. coli cells match with the open reading frame of the ung gene. The protein sequence analysis shows that the N-terminal methionine is cleaved off in the mature protein. The in vitro transcription coupled translation of the ung gene directs the synthesis of a protein which comigrates with uracil DNA glycosylase. Also, the CNBr cleavage of the protein synthesized in vitro confirms the positions of the methionines deduced from the DNA sequence. The levels of ung gene expression remain constant up to the early stationary phase, but decline in the late stationary phase of the E. coli culture. The E. coli gene showed a strong sequence homology to Shigella, a weak sequence homology to Salmonella and Citrobacter, and a very weak sequence homology to Proteus genes. No sequence homologies were seen for Pseudomonas, Clostridium, Micrococcus, and several eukaryotic genomes.
Varshney
U
U
Department of Medical Biochemistry, Faculty of Medicine, University of Calgary, Canada.
Hutcheon
T
T
van de Sande
J H
JH
D000595:Amino Acid Sequence; D001483:Base Sequence; D045647:DNA Glycosylases; D004720:Endonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000032:analysis / Q000235:genetics*; D010449:Peptide Mapping; D014176:Protein Biosynthesis; D012260:Ribonucleases / Q000378:metabolism; D015719:Single-Strand Specific DNA and RNA Endonucleases; D014158:Transcription, Genetic; D051981:Uracil-DNA Glycosidase
D004720:Endonucleases; D012260:Ribonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase