Role for the leucine-responsive regulatory protein (Lrp) as a structural protein in regulating the Escherichia coli gcvTHP operon.
The Escherichia coli glycine-cleavage enzyme system (gcvTHP and lpd gene products) provides C1 units for cellular methylation reactions. Both the GcvA and leucine-responsive regulatory (Lrp) proteins are required for regulation of the gcv operon. One model proposed for gcv regulation is that Lrp plays a structural role, bending the DNA to allow GcvA to function as either an activator or a repressor in response to environmental signals. This hypothesis was tested by replacing all but the upstream 22 bp of the Lrp-binding region in a gcvT::lacZ fusion with the I1A site from phage lambda. Integration host factor (IHF) binds the I1A site and bends the DNA about 140 degrees. Shifting the I1A site by increments of 1 base around the DNA helix resulted in IHF-dependent activation and repression of gcvT::lacZ expression that were face-of-the-helix dependent. Activation was also dependent on the GcvA protein, and repression was dependent on both the GcvA and GcvR proteins, demonstrating that the roles for these proteins were not altered. The results are consistent with Lrp playing primarily a structural role in gcv regulation, although they do not completely rule out the possibility that Lrp also interacts with another gcv-regulatory protein or with RNA polymerase.
Stauffer
Lorraine T
LT
Stauffer
George V
GV
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D050960:Aminomethyltransferase; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004789:Enzyme Activation; D004794:Enzyme Repression; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D005998:Glycine / Q000378:metabolism*; D019877:Hydroxymethyl and Formyl Transferases / Q000096:biosynthesis / Q000235:genetics; D034982:Integration Host Factors; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D009097:Multienzyme Complexes / Q000096:biosynthesis / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009876:Operon; D011485:Protein Binding; D011971:Receptors, Immunologic / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014166:Transferases / Q000096:biosynthesis / Q000235:genetics*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C095454:GcvA protein, E coli; D034982:Integration Host Factors; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D009097:Multienzyme Complexes; D011971:Receptors, Immunologic; D014157:Transcription Factors; C064525:glycine cleavage system; D000594:Amino Acid Oxidoreductases; C087190:GcvP protein, E coli; D014166:Transferases; D019877:Hydroxymethyl and Formyl Transferases; D050960:Aminomethyltransferase; D005998:Glycine