Identification of a Functional Nuclear Localization Signal Mediating Nuclear Import of the Zinc Finger Transcription Factor ZNF24

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Nov 14

PMID 24224020

Citations 7

Authors

Jian-Zhong Li

Xia Chen

Xue-Lian Gong

Hong-Yuan Hu

Duo Shi

Yi-Ming Lu

Lei Qiu

Fa Lu

Zhen-Lin Hu

Jun-Ping Zhang

Affiliations

Soon will be listed here.

Abstract

ZNF24 is a member of the SCAN domain family of Krüppel-like zinc finger (ZF) transcription factors, which plays a critical role in cell proliferation and differentiation. However, how ZNF24 enters the nucleus in order to exert its function remains unclear since its nuclear localization signal(s) (NLS) has not been identified. Here, we generated a series of GFP-tagged deletion and point mutants and assessed their subcellular localization. Our results delimit the NLS to ZF1-2. Deletion of ZF1-2 caused cytoplasmic accumulation of ZNF24. Fusion of the ZF1-2 to green fluorescent protein (GFP) targeted GFP to the nucleus, demonstrating that the ZF1-2 is both necessary and sufficient for nuclear localization. ZNF24 containing histidine to leucine mutations that disrupt the structure of ZF1 or/and ZF2 retains appropriate nuclear localization, indicating that neither the tertiary structure of the zinc fingers nor specific DNA binding are necessary for nuclear localization. K286A and R290A mutation led to partial cytoplasmic accumulation. Co-immunoprecipitation demonstrated that ZNF24 interacted with importin-β and this interaction required the ZF motifs. The β-Catenin (CTNNB1) luciferase assays showed that the ZNF24 mutants defective in nuclear localization could not promote CTNNB1 promoter activation as the wild-type ZNF24 did. Taken together, these results suggest that consecutive ZF1-2 is critical for the regulation of ZNF24 nuclear localization and its transactivation function.

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References

Li J, Chen X, Liu Y, Ding L, Qiu L, Hu Z . The transcriptional repression of platelet-derived growth factor receptor-beta by the zinc finger transcription factor ZNF24. Biochem Biophys Res Commun. 2010; 397(2):318-22. DOI: 10.1016/j.bbrc.2010.05.110. View

Kuwahara J, Watanabe Y, Kayasuga T, Itoh K . Zn finger and nuclear localization of transcription factor Sp1. Nucleic Acids Symp Ser. 2003; (44):265-6. DOI: 10.1093/nass/44.1.265. View

Takei Y, Yamamoto K, Tsujimoto G . Identification of the sequence responsible for the nuclear localization of human Cdc6. FEBS Lett. 1999; 447(2-3):292-6. DOI: 10.1016/s0014-5793(99)00306-3. View

Rousseau-Merck M, Huebner K, Berger R, Thiesen H . Chromosomal localization of two human zinc finger protein genes, ZNF24 (KOX17) and ZNF29 (KOX26), to 18q12 and 17p13-p12, respectively. Genomics. 1991; 9(1):154-61. DOI: 10.1016/0888-7543(91)90233-5. View

Hatayama M, Tomizawa T, Sakai-Kato K, Bouvagnet P, Kose S, Imamoto N . Functional and structural basis of the nuclear localization signal in the ZIC3 zinc finger domain. Hum Mol Genet. 2008; 17(22):3459-73. PMC: 2572694. DOI: 10.1093/hmg/ddn239. View

Michalowski C, Pfanzagl B, Loffelhardt W, Bohnert H . The cyanelle S10 spc ribosomal protein gene operon from Cyanophora paradoxa. Mol Gen Genet. 1990; 224(2):222-31. DOI: 10.1007/BF00271555. View

Nam K, Honer C, Schumacher C . Structural components of SCAN-domain dimerizations. Proteins. 2004; 56(4):685-92. DOI: 10.1002/prot.20170. View

Klug A, Schwabe J . Protein motifs 5. Zinc fingers. FASEB J. 1995; 9(8):597-604. View

Harper J, Yan L, Loureiro R, Wu I, Fang J, DAmore P . Repression of vascular endothelial growth factor expression by the zinc finger transcription factor ZNF24. Cancer Res. 2007; 67(18):8736-41. DOI: 10.1158/0008-5472.CAN-07-1617. View

10.

Mehta T, Lu H, Wang X, Urvalek A, Nguyen K, Monzur F . A unique sequence in the N-terminal regulatory region controls the nuclear localization of KLF8 by cooperating with the C-terminal zinc-fingers. Cell Res. 2009; 19(9):1098-109. DOI: 10.1038/cr.2009.64. View

11.

Wali A, Ali G, John P, Lee K, Chishti M, Leal S . Mapping of a gene for alopecia with mental retardation syndrome (APMR3) on chromosome 18q11.2-q12.2. Ann Hum Genet. 2007; 71(Pt 5):570-7. PMC: 6148758. DOI: 10.1111/j.1469-1809.2007.00362.x. View

12.

Zhao J . Sumoylation regulates diverse biological processes. Cell Mol Life Sci. 2007; 64(23):3017-33. PMC: 7079795. DOI: 10.1007/s00018-007-7137-4. View

13.

Khalfallah O, Ravassard P, Lagache C, Fligny C, Serre A, Bayard E . Zinc finger protein 191 (ZNF191/Zfp191) is necessary to maintain neural cells as cycling progenitors. Stem Cells. 2009; 27(7):1643-53. DOI: 10.1002/stem.88. View

14.

Han Z, Zhang Q, Ye M, Kan L, Gu B, He K . Molecular cloning of six novel Krüppel-like zinc finger genes from hematopoietic cells and identification of a novel transregulatory domain KRNB. J Biol Chem. 1999; 274(50):35741-8. DOI: 10.1074/jbc.274.50.35741. View

15.

Weis K . Regulating access to the genome: nucleocytoplasmic transport throughout the cell cycle. Cell. 2003; 112(4):441-51. DOI: 10.1016/s0092-8674(03)00082-5. View

16.

Corbi N, Libri V, Onori A, Passananti C . Synthetic zinc finger peptides: old and novel applications. Biochem Cell Biol. 2004; 82(4):428-36. DOI: 10.1139/o04-047. View

17.

Shi S, Liu M, Yu L, Chen S, Zheng Q, Wu G . [Assignment of a novel zinc finger gene ZNF191 to human chromosome 18Q12.1 by human/rodent somatic cell hybrid panel and fluorescent in situ hybridization]. Shi Yan Sheng Wu Xue Bao. 2002; 31(1):21-7. View

18.

Sander G, Marsh R, Parmeggiani A . Activity of the 30-S CsCl core in elongation-factor-dependent GTP hydrolysis. Eur J Biochem. 1976; 61(1):317-23. DOI: 10.1111/j.1432-1033.1976.tb10024.x. View

19.

Edelstein L, Collins T . The SCAN domain family of zinc finger transcription factors. Gene. 2005; 359:1-17. DOI: 10.1016/j.gene.2005.06.022. View

20.

Nedved M, Moe G . Cooperative, non-specific binding of a zinc finger peptide to DNA. Nucleic Acids Res. 1994; 22(22):4705-11. PMC: 308521. DOI: 10.1093/nar/22.22.4705. View