Structural approach to a novel tandem repeat DNA-binding domain, STPR, by CD and NMR

Biochemistry. 2007 Feb 20;46(7):1703-13. doi: 10.1021/bi061780c. Epub 2007 Jan 24.

Abstract

Fibroin-modulator-binding protein 1 (FMBP-1) is a factor that binds the transcriptional activation elements of the fibroin gene. It has a novel structure, consisting of four tandem repeats (R1-R4) of 23 amino acids each in the C-terminal half. This region is referred to as the STPR (score and three amino acid peptide repeat) domain and acts as a DNA-binding domain in FMBP-1. Interestingly, the homology among the four repeats is remarkably high. Here, we have determined the three-dimensional structures of the four repeats by NMR. All four repeat units have basically the same structure: a short alpha-helix in the N-terminal half maintained by a salt bridge and an N-capping box. CD studies showed that the full-length STPR domain was 31% helical in solution. This is explained by the connections among the four short helices that were determined separately by NMR. From the thermal-denaturation study, it can be deduced that these four helices in the full-length STPR domain moved flexibly with no interaction among them. However, the specific DNA caused a distinct increase, of up to 76%, in the alpha-helical content of the full-length STPR domain. This finding suggests that the binding of the full-length STPR domain to specific DNA causes an induced-fit conformational change that increases alpha-helicity; the poorly structured regions of the protein may form a regular secondary structure. Furthermore, the mutation analysis showed that the four repeats of the STPR domain raise the possibility of interaction with DNA in different ways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bombyx
  • Circular Dichroism
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics
  • Fibroins / genetics*
  • Insect Proteins / chemistry*
  • Insect Proteins / genetics
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Denaturation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Repetitive Sequences, Amino Acid
  • Sequence Homology, Amino Acid
  • Solutions
  • Transcription Factors / chemistry*
  • Transcription Factors / genetics

Substances

  • DNA-Binding Proteins
  • Insect Proteins
  • Solutions
  • Transcription Factors
  • Fibroins

Associated data

  • PDB/1VD7
  • PDB/1VD8
  • PDB/1VD9
  • PDB/1VDA