A peptide C-terminal to the second Zn finger of human vitamin D receptor is able to specify nuclear localization

Eur J Biochem. 1994 Jul 15;223(2):381-7. doi: 10.1111/j.1432-1033.1994.tb19004.x.

Abstract

A peptide of 27 amino acids, VDR(102-76), representing residues 76-102 immediately C-terminal to the second Zn finger of human vitamin D receptor (hVDR) was conjugated to fluorescein-labelled IgG using a bifunctional coupling reagent, m-maleimidobenzoyl n-hydroxysuccinimide. Upon microinjection into the cytoplasm of human osteosarcoma MG-63 cells, the chimeras accumulated in the nuclei. This transport was arrested by chilling or energy depletion. Two other peptides, VDR(80-67), spanning the N-terminal part of VDR(102-76), and VDR(108-97), spanning the C-terminal part of VDR(102-76), were not able to target the linked proteins to the nuclei. SV40(135-112), a peptide containing a well-characterized nuclear localization sequence (amino acids 112-135) of simian virus 40 (SV40) large T-antigen, caused complete nuclear accumulation under the same conditions. Wheat germ agglutinin, which inhibits SV40(135-112) transport, also inhibited the nuclear accumulation of VDR(102-76) as did energy depletion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antigens, Polyomavirus Transforming / chemistry
  • Antigens, Polyomavirus Transforming / metabolism
  • Biological Transport
  • Cell Nucleus / metabolism*
  • Cold Temperature
  • Cross-Linking Reagents
  • Electrophoresis, Polyacrylamide Gel
  • Energy Metabolism
  • Fluorescein-5-isothiocyanate
  • Humans
  • Immunoglobulin G
  • Molecular Sequence Data
  • Osteosarcoma
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism*
  • Receptors, Calcitriol / chemistry
  • Receptors, Calcitriol / metabolism*
  • Tumor Cells, Cultured
  • Wheat Germ Agglutinins / pharmacology
  • Zinc Fingers*

Substances

  • Antigens, Polyomavirus Transforming
  • Cross-Linking Reagents
  • Immunoglobulin G
  • Peptide Fragments
  • Receptors, Calcitriol
  • Wheat Germ Agglutinins
  • Fluorescein-5-isothiocyanate