Nucleic Acids Research, 2000, Vol. 28, No. 12 2389-2395
© 2000 Oxford University Press
DNA-binding sequence of the human prostate-specific homeodomain protein NKX3.1
Department of Oncology, Lombardi Cancer Center, Georgetown University School of Medicine, 3800 Reservoir Road NW, Washington, DC 20007-2197, USA
NKX3.1 is a member of the NK class of homeodomain proteins and is most closely related to Drosophila NK-3. NKX3.1 has predominantly prostate-specific expression in the adult human. Previous studies suggested that NKX3.1 exerts a growth-suppressive effect on prostatic epithelial cells and controls differentiated glandular functions. Using a binding site selection assay with recombinant NKX3.1 protein we identified a TAAGTA consensus binding sequence that has not been reported for any other NK class homeoprotein. By electromobility shift assay we demonstrated that NKX3.1 preferentially binds the TAAGTA sequence rather than the binding site for Nkx2.1 (CAAGTG) or Msx1 (TAATTG). Using mutated binding sites in competitive gel shift assays, we analyzed the nucleotides in the TAAGTA consensus sequence that are important for NKX3.1 binding. The consensus binding site of a naturally occurring polymorphic NKX3.1 protein with arginine replaced by cysteine at position 52 was identical to the wild-type binding sequence. The binding affinities of wild-type and polymorphic NKX3.1 for the TAAGTA consensus site were very similar, with values of 20 and 22 nM, respectively. Wild-type and polymorphic NKX3.1 specifically repressed transcription of luciferase from a reporter vector with three copies of the NKX3.1-binding site upstream from a thymidine kinase promoter. The data show that among NK family proteins NKX3.1 binds a novel DNA sequence and can behave as an in vitro transcriptional repressor.
* To whom correspondence should be addressed. Tel: +1 202 687 2207; Fax: +1 202 784 1229; Email: gelmanne@gunet.georgetown.edu
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  K. A. Holmes, J. S. Song, X. S. Liu, M. Brown, and J. S. Carroll Nkx3-1 and LEF-1 Function as Transcriptional Inhibitors of Estrogen Receptor Activity Cancer Res., September 15, 2008; 68(18): 7380 - 7385. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Markowski, C. Bowen, and E. P. Gelmann Inflammatory Cytokines Induce Phosphorylation and Ubiquitination of Prostate Suppressor Protein NKX3.1 Cancer Res., September 1, 2008; 68(17): 6896 - 6901. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. P. Mogal, R. van der Meer, P. S. Crooke, and S. A. Abdulkadir Haploinsufficient Prostate Tumor Suppression by Nkx3.1: A ROLE FOR CHROMATIN ACCESSIBILITY IN DOSAGE-SENSITIVE GENE REGULATION J. Biol. Chem., August 31, 2007; 282(35): 25790 - 25800. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Bowen, A. Stuart, J.-H. Ju, J. Tuan, J. Blonder, T. P. Conrads, T. D. Veenstra, and E. P. Gelmann NKX3.1 Homeodomain Protein Binds to Topoisomerase I and Enhances Its Activity Cancer Res., January 15, 2007; 67(2): 455 - 464. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Choi and A. Rajkovic Characterization of NOBOX DNA Binding Specificity and Its Regulation of Gdf9 and Pou5f1 Promoters J. Biol. Chem., November 24, 2006; 281(47): 35747 - 35756. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Zheng, J.-h. Ju, B.-l. Chang, E. Ortner, J. Sun, S. D. Isaacs, J. Sun, K. E. Wiley, W. Liu, M. Zemedkun, et al. Germ-Line Mutation of NKX3.1 Cosegregates with Hereditary Prostate Cancer and Alters the Homeodomain Structure and Function Cancer Res., January 1, 2006; 66(1): 69 - 77. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Asatiani, W.-X. Huang, A. Wang, E. Rodriguez Ortner, L. R. Cavalli, B. R. Haddad, and E. P. Gelmann Deletion, Methylation, and Expression of the NKX3.1 Suppressor Gene in Primary Human Prostate Cancer Cancer Res., February 15, 2005; 65(4): 1164 - 1173. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Hosohata, P. Li, Y. Hosohata, J. Qin, R. G. Roeder, and Z. Wang Purification and Identification of a Novel Complex Which Is Involved in Androgen Receptor-Dependent Transcription Mol. Cell. Biol., October 1, 2003; 23(19): 7019 - 7029. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W.-S. Cheng, V. Giandomenico, I. Pastan, and M. Essand Characterization of the Androgen-Regulated Prostate-Specific T Cell Receptor {gamma}-Chain Alternate Reading Frame Protein (TARP) Promoter Endocrinology, August 1, 2003; 144(8): 3433 - 3440. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. G. Nelson, A. M. De Marzo, and W. B. Isaacs Prostate Cancer N. Engl. J. Med., July 24, 2003; 349(4): 366 - 381. [Full Text] [PDF]
|
|
|
|
|
|
D.-W. Kim, H. Kempf, R. E. Chen, and A. B. Lassar Characterization of Nkx3.2 DNA Binding Specificity and Its Requirement for Somitic Chondrogenesis J. Biol. Chem., July 18, 2003; 278(30): 27532 - 27539. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L.-H. Wang, R. Chmelik, and M. Nirenberg Sequence-specific DNA binding by the vnd/NK-2 homeodomain of Drosophila PNAS, October 1, 2002; 99(20): 12721 - 12726. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. P. Gelmann, D. J. Steadman, J. Ma, N. Ahronovitz, H. J. Voeller, S. Swope, M. Abbaszadegan, K. M. Brown, K. Strand, R. B. Hayes, et al. Occurrence of NKX3.1 C154T Polymorphism in Men with and without Prostate Cancer and Studies of Its Effect on Protein Function Cancer Res., May 1, 2002; 62(9): 2654 - 2659. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Chen, A. K. Nandi, X. Li, and C. J. Bieberich NKX-3.1 Interacts with Prostate-derived Ets Factor and Regulates the Activity of the PSA Promoter Cancer Res., January 1, 2002; 62(2): 338 - 340. [Abstract] [Full Text] [PDF]
|
|