A peptide C-terminal to the second Zn finger of human vitamin D receptor is able to specify nuclear localization.
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.
Luo
Z
Z
Department of Biochemistry & Biotechnology, University of Kuopio, Finland.
Rouvinen
J
J
Mäenpää
P H
PH
D000595:Amino Acid Sequence; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000378:metabolism; D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism*; D003080:Cold Temperature; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D007074:Immunoglobulin G; D008969:Molecular Sequence Data; D012516:Osteosarcoma; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D018167:Receptors, Calcitriol / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014909:Wheat Germ Agglutinins / Q000494:pharmacology; D016335:Zinc Fingers
D000952:Antigens, Polyomavirus Transforming; D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D010446:Peptide Fragments; D018167:Receptors, Calcitriol; D014909:Wheat Germ Agglutinins; D016650:Fluorescein-5-isothiocyanate