In vitro selection of high affinity HspR-binding sites within the genome of Helicobacter pylori.
The major chaperone genes of Helicobacter pylori are negatively regulated by HspR, a homologue of the repressor of the dnaK operon of Streptomyces coelicolor. Using an in vitro selection and amplification approach we identified two new chromosomal binding sites of the HspR protein. Both binding sites were characterized by footprinting analysis with purified HspR protein. Intriguingly, these HspR binding sites are located at the 3prime prime or minute ends of two genes coding for predicted proteins with functions unrelated to those of chaperones. This suggests that H. pylori HspR may regulate the expression of genes encoding proteins with diverse functions. Nucleotide sequence alignment of HspR-binding sites highlights conserved nucleotides extending outside the previously proposed consensus binding sequence with structural features predicting geometry of HspR binding as an oligomer.
Delany
Isabel
I
Department of Molecular Biology, IRIS-Chiron S.p.A., Via Fiorentina 1, 53100, Siena, Italy.
Spohn
Gunther
G
Rappuoli
Rino
R
Scarlato
Vincenzo
V
D001426:Bacterial Proteins; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018983:DNA Footprinting; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D016680:Genome, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D016480:Helicobacter pylori / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins; C108444:HspR protein, bacteria; D012097:Repressor Proteins; D003850:Deoxyribonuclease I