Characterization and structure of the manganese-responsive transcriptional regulator ScaR

Biochemistry. 2009 Nov 3;48(43):10308-20. doi: 10.1021/bi900980g.

Abstract

The streptococcal coaggregation regulator (ScaR) of Streptococcus gordonii is a manganese-dependent transcriptional regulator. When intracellular manganese concentrations become elevated, ScaR represses transcription of the scaCBA operon, which encodes a manganese uptake transporter. A member of the DtxR/MntR family of metalloregulators, ScaR shares sequence similarity with other family members, and many metal-binding residues are conserved. Here, we show that ScaR is an active dimer, with two dimers binding the 46 base pair scaC operator. Each ScaR subunit binds two manganese ions, and the protein is activated by a variety of other metal ions, including Cd(2+), Co(2+), and Ni(2+) but not Zn(2+). The crystal structure of apo-ScaR reveals a tertiary and quaternary structure similar to its homologue, the iron-responsive regulator DtxR. While each DtxR subunit binds a metal ion in two sites, labeled primary and ancillary, crystal structures of ScaR determined in the presence of Cd(2+) and Zn(2+) show only a single occupied metal-binding site that is novel to ScaR. The site analogous to the primary site in DtxR is unoccupied, and the ancillary site is absent from ScaR. Instead, metal ions bind to ScaR at a site labeled "secondary", which is composed of Glu80, Cys123, His125, and Asp160 and lies roughly 5 A away from where the ancillary site would be predicted to exist. This difference suggests that ScaR and its closely related homologues are activated by a mechanism distinct from that of either DtxR or MntR.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Cadmium / pharmacology
  • Chromatography, Gel
  • Cobalt / pharmacology
  • Crystallography, X-Ray
  • DNA / metabolism
  • Gene Expression Regulation, Bacterial / drug effects
  • Gene Expression Regulation, Bacterial / genetics
  • Manganese / pharmacology*
  • Molecular Sequence Data
  • Nickel / pharmacology
  • Protein Binding / drug effects
  • Protein Multimerization
  • Sequence Homology, Amino Acid
  • Streptococcus / metabolism*
  • Ultracentrifugation
  • Zinc / pharmacology

Substances

  • Bacterial Proteins
  • Cadmium
  • Cobalt
  • Manganese
  • Nickel
  • DNA
  • Zinc

Associated data

  • PDB/3HRS
  • PDB/3HRT
  • PDB/3HRU