A universal stress protein (USP) in mycobacteria binds cAMP

J Biol Chem. 2015 May 15;290(20):12731-43. doi: 10.1074/jbc.M115.644856. Epub 2015 Mar 23.


Mycobacteria are endowed with rich and diverse machinery for the synthesis, utilization, and degradation of cAMP. The actions of cyclic nucleotides are generally mediated by binding of cAMP to conserved and well characterized cyclic nucleotide binding domains or structurally distinct cGMP-specific and -regulated cyclic nucleotide phosphodiesterase, adenylyl cyclase, and E. coli transcription factor FhlA (GAF) domain-containing proteins. Proteins with cyclic nucleotide binding and GAF domains can be identified in the genome of mycobacterial species, and some of them have been characterized. Here, we show that a significant fraction of intracellular cAMP is bound to protein in mycobacterial species, and by using affinity chromatography techniques, we identify specific universal stress proteins (USP) as abundantly expressed cAMP-binding proteins in slow growing as well as fast growing mycobacteria. We have characterized the biochemical and thermodynamic parameters for binding of cAMP, and we show that these USPs bind cAMP with a higher affinity than ATP, an established ligand for other USPs. We determined the structure of the USP MSMEG_3811 bound to cAMP, and we confirmed through structure-guided mutagenesis, the residues important for cAMP binding. This family of USPs is conserved in all mycobacteria, and we suggest that they serve as "sinks" for cAMP, making this second messenger available for downstream effectors as and when ATP levels are altered in the cell.

Keywords: Crystal Structure; Cyclic AMP (cAMP); Isothermal Titration Calorimetry (ITC); Mycobacteria; Surface Plasmon Resonance (SPR); Universal Stress Protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate* / genetics
  • Adenosine Triphosphate* / metabolism
  • Bacterial Proteins* / chemistry
  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • Cyclic AMP* / chemistry
  • Cyclic AMP* / metabolism
  • Genome, Bacterial
  • Heat-Shock Proteins* / chemistry
  • Heat-Shock Proteins* / genetics
  • Heat-Shock Proteins* / metabolism
  • Mycobacterium* / chemistry
  • Mycobacterium* / genetics
  • Mycobacterium* / metabolism
  • Second Messenger Systems / physiology*


  • Bacterial Proteins
  • Heat-Shock Proteins
  • Adenosine Triphosphate
  • Cyclic AMP

Associated data

  • PDB/1MJH
  • PDB/1TQ8
  • PDB/2BA
  • PDB/2Z08
  • PDB/5ahw