The central domain of Escherichia coli TyrR is responsible for hexamerization associated with tyrosine-mediated repression of gene expression.
TyrR from Escherichia coli regulates the expression of genes for aromatic amino acid uptake and biosynthesis. Its central ATP-hydrolyzing domain is similar to conserved domains of bacterial regulatory proteins that interact with RNA polymerase holoenzyme associated with the alternative sigma factor, sigma(54). It is also related to the common module of the AAA+ superfamily of proteins that is involved in a wide range of cellular activities. We expressed and purified two TyrR central domain polypeptides. The fragment comprising residues 188-467, called TyrR-(188-467), was soluble and stable, in contrast to that corresponding to the conserved core from residues 193 to 433. TyrR-(188-467) bound ATP and rhodamine-ATP with association constants 2- to 5-fold lower than TyrR and hydrolyzed ATP at five times the rate of TyrR. In contrast to TyrR, which is predominantly dimeric at protein concentrations less than 10 microm in the absence of ligands, or in the presence of ATP or tyrosine alone, TyrR-(188-467) is a monomer, even at high protein concentrations. Tyrosine in the presence of ATP or ATPgammaS promotes the oligomerization of TyrR-(188-467) to a hexamer. Tyrosine-dependent repression of gene transcription by TyrR therefore depends on ligand binding and hexamerization determinants located in the central domain polypeptide TyrR-(188-467).
Dixon
Mathew P
MP
Department of Biochemistry and Molecular Biology, The University of Melbourne, Parkville 3010, Australia.
Pau
Richard N
RN
Howlett
Geoffrey J
GJ
Dunstan
David E
DE
Sawyer
William H
WH
Davidson
Barrie E
BE
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000737:chemistry* / Q000494:pharmacology* / Q000502:physiology; D012235:Rhodamines / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism; D014443:Tyrosine / Q000494:pharmacology*
D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D012235:Rhodamines; C069256:TyrR protein, E coli; D014443:Tyrosine; D014364:Tryptophan; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases