Identification of bZIP interaction partners of viral proteins HBZ, MEQ, BZLF1, and K-bZIP using coiled-coil arrays

Biochemistry. 2010 Mar 9;49(9):1985-97. doi: 10.1021/bi902065k.

Abstract

Basic-region leucine-zipper transcription factors (bZIPs) contain a segment rich in basic amino acids that can bind DNA, followed by a leucine zipper that can interact with other leucine zippers to form coiled-coil homo- or heterodimers. Several viruses encode proteins containing bZIP domains, including four that encode bZIPs lacking significant homology to any human protein. We investigated the interaction specificity of these four viral bZIPs by using coiled-coil arrays to assess self-associations as well as heterointeractions with 33 representative human bZIPs. The arrays recapitulated reported viral-human interactions and also uncovered new associations. MEQ and HBZ interacted with multiple human partners and had unique interaction profiles compared to any human bZIPs, whereas K-bZIP and BZLF1 displayed homospecificity. New interactions detected included HBZ with MAFB, MAFG, ATF2, CEBPG, and CREBZF and MEQ with NFIL3. These were confirmed in solution using circular dichroism. HBZ can heteroassociate with MAFB and MAFG in the presence of MARE-site DNA, and this interaction is dependent on the basic region of HBZ. NFIL3 and MEQ have different yet overlapping DNA-binding specificities and can form a heterocomplex with DNA. Computational design considering both affinity for MEQ and specificity with respect to other undesired bZIP-type interactions was used to generate a MEQ dimerization inhibitor. This peptide, anti-MEQ, bound MEQ both stably and specifically, as assayed using coiled-coil arrays and circular dichroism in solution. Anti-MEQ also inhibited MEQ binding to DNA. These studies can guide further investigation of the function of viral and human bZIP complexes.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Avian Proteins / chemistry
  • Avian Proteins / metabolism
  • Basic-Leucine Zipper Transcription Factors / chemistry
  • Basic-Leucine Zipper Transcription Factors / metabolism*
  • Chickens
  • Genome, Viral
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism*
  • Oncogene Proteins, Viral / chemistry
  • Oncogene Proteins, Viral / metabolism*
  • Protein Array Analysis*
  • Protein Structure, Tertiary
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / metabolism*
  • Repressor Proteins / chemistry
  • Repressor Proteins / metabolism*
  • Reproducibility of Results
  • Retroviridae Proteins
  • Trans-Activators / chemistry
  • Trans-Activators / metabolism*
  • Transcription Factors / chemistry
  • Transcription Factors / metabolism*
  • Viral Proteins / chemistry
  • Viral Proteins / metabolism*

Substances

  • ALYREF protein, human
  • Avian Proteins
  • BZLF1 protein, Herpesvirus 4, Human
  • Basic-Leucine Zipper Transcription Factors
  • Eco-Q protein, Gallid herpesvirus 2
  • HBZ protein, human T-cell leukemia virus type I
  • K8 protein, Human herpesvirus 8
  • Nuclear Proteins
  • Oncogene Proteins, Viral
  • RNA-Binding Proteins
  • Repressor Proteins
  • Retroviridae Proteins
  • Trans-Activators
  • Transcription Factors
  • Viral Proteins