Novel nuclear localization signal between the two DNA-binding zinc fingers in the human vitamin D receptor

Jui Cheng Hsieh, Yoshiko Shimizu, Shinsei Minoshima, Nobuyoshi Shimizu, Carol A. Haussler, Peter Jurutka, Mark R. Haussler

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The human vitamin D receptor (hVDR) possesses a unique array of five basic amino acids positioned between the two DNA-binding zinc fingers that is similar to well-characterized nuclear localization sequences in other proteins. When residues within this region are mutated to nonbasic amino acids, or when this domain is deleted, the receptor is still well expressed, but it no longer associates with the vitamin D-responsive element in DNA, in vitro, and hVDR-mediated transcriptional activation is abolished in transfected cells. Concomitantly, the mutated hVDRs exhibit a significant shift in hVDR cellular distribution favoring cytoplasmic over nuclear retention as assessed by subcellular fractionation and immunoblotting. Independent immunocytochemical studies employing a VDR-specific monoclonal antibody demonstrate that mutation or deletion of this basic domain dramatically attenuates hVDR nuclear localization in transfected COS-7 cells. Although wild-type hVDR is partitioned predominantly to the nucleus in the absence of the 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) hormone, treatment with ligand further enhances nuclear translocation, as it does to some degree in receptors with the basic region altered. The role of 1,25(OH)2D3 may be to facilitate hVDR heterodimerization with retinoid X receptors, stimulating subsequent DNA binding and ultimately enhancing nuclear retention. Taken together, these data reveal that the region of hVDR between Arg-49 and Lys-55 contains a novel constitutive nuclear localization signal, RRSMKRK.

Original languageEnglish (US)
Pages (from-to)94-109
Number of pages16
JournalJournal of Cellular Biochemistry
Volume70
Issue number1
DOIs
StatePublished - Jul 1 1998
Externally publishedYes

Fingerprint

Nuclear Localization Signals
Calcitriol Receptors
Zinc Fingers
Zinc
DNA
Retinoid X Receptors
Basic Amino Acids
Calcitriol
Sequence Deletion
COS Cells
Fractionation
Immunoblotting
Vitamin D
Transcriptional Activation
Chemical activation
Monoclonal Antibodies
Hormones
Ligands
Amino Acids

Keywords

  • 1,25-dihydroxyvitamin D
  • DNA-binding
  • Nuclear retention
  • Steroid hormone receptor
  • Transcriptional activation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Novel nuclear localization signal between the two DNA-binding zinc fingers in the human vitamin D receptor. / Hsieh, Jui Cheng; Shimizu, Yoshiko; Minoshima, Shinsei; Shimizu, Nobuyoshi; Haussler, Carol A.; Jurutka, Peter; Haussler, Mark R.

In: Journal of Cellular Biochemistry, Vol. 70, No. 1, 01.07.1998, p. 94-109.

Research output: Contribution to journalArticle

Hsieh, Jui Cheng ; Shimizu, Yoshiko ; Minoshima, Shinsei ; Shimizu, Nobuyoshi ; Haussler, Carol A. ; Jurutka, Peter ; Haussler, Mark R. / Novel nuclear localization signal between the two DNA-binding zinc fingers in the human vitamin D receptor. In: Journal of Cellular Biochemistry. 1998 ; Vol. 70, No. 1. pp. 94-109.
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