title: Neglected role of cAMP receptor protein monomer.
abstract: Lac operon transcription activation through CRP dimer depends on cAMP second messenger. The formation of CRP homodimers is mediated by protein-protein interactions between the monomers. Cyclic AMP ligand binding brings CRP dimer to an active state via conformational changes. Molecular modeling studies in our lab showed the importance of monomer in transcription activation through its pre-existing conformational state. Until now CRP experiments were carried out at protein concentrations higher than that of CRP dimer dissociation value making all CRP monomers dimer and ignore the importance of CRP monomer in allosteric activation. Labeling CRP monomers with fluorophores exterminate using excess protein concentration and allow monitoring CRP monomer behavior. CRP monomer exchange accelerates in the presence of non specific DNA whereas the exchange is inhibited in the presence of specific DNA and cAMP ligand. Degree of subunit exchange depends on the stability of CRP dimer. Cyclic AMP forms a single molecule from two monomers and addition of specific DNA further stabilizes CRP dimer and decreases monomer exchange. On the other hand, addition of non specific DNA increases CRP monomer exchange and may explain the mechanism of CRP monomer removal and dissociation of CRP dimer:cAMP:DNA complex. The exchange behavior of CRP in the presence of different factors implies importance of monomer in transcription complex association and dissociation.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Cumhuriyet University, 58140, Sivas, Turkey. ytutar@cumhuriyet.edu.tr
authors.forename: Yusuf
authors.initials: Y
authors.lastname: Tutar
mesh_terms: D000818:Animals; D002417:Cattle; D000242:Cyclic AMP / Q000378:metabolism; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D055503:Protein Multimerization; D017433:Protein Structure, Secondary
chemical_list: D002373:Cyclic AMP Receptor Protein; D004247:DNA; C427203:calf thymus DNA; D000242:Cyclic AMP