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Molecular and Cellular Biology, March 2006, p. 2387-2398, Vol. 26, No. 6
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.6.2387-2398.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Regulation of PCNA and Cyclin D1 Expression and Epithelial Morphogenesis by the ZO-1-Regulated Transcription Factor ZONAB/DbpA{dagger}

Tony Sourisseau,1 Anastasios Georgiadis,2 Anna Tsapara,1 Robin R. Ali,2 Richard Pestell,3 Karl Matter,1* and Maria S. Balda1*

Divisions of Cell Biology,1 Molecular Therapy,2 Institute of Ophthalmology, University College London, London, United Kingdom, and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania3

Received 14 June 2005/ Returned for modification 5 August 2005/ Accepted 3 January 2006

The tight junction protein ZO-1 inhibits G1/S-phase transition by cytoplasmic sequestration of a complex formed by CDK4 and the transcription factor ZONAB. Canine ZONAB is the homologue of human DbpA, an E2F target gene that is overexpressed in different carcinomas. Since the ZONAB target genes that are involved in G1/S-phase transition are unknown, we employed the mammary epithelial cell line MCF-10A and cDNA arrays to screen for such genes. We identified genes encoding cell cycle and replication proteins whose expression was altered due to increased ZONAB expression. For proliferative cell nuclear antigen and cyclin D1 genes, we show that increased mRNA levels resulted in increased protein levels and we identified ZONAB-responsive elements in their promoters by using different approaches, including chromatin immunoprecipitation assays. RNA interference and overexpression of ZONAB affected the proliferation of both MCF-10A and MDCK cells as well as the differentiation of MDCK cells into polarized cysts in three-dimensional cultures. These results indicate that ZONAB regulates the transcription of genes that are important for G1/S-phase progression and links tight junctions to the transcriptional control of key cell cycle regulators and epithelial cell differentiation.

* Corresponding author. Mailing address: Division of Cell Biology, Institute of Ophthalmology, University College London, Bath Street, London EC1V 9EL, United Kingdom. Phone: 0044 20 7608 6861/4014. Fax: 0044 20 7608 4034. E-mail for Maria S. Balda: m.balda{at}ucl.ac.uk. E-mail for Karl Matter: k.matter{at}ucl.ac.uk.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, March 2006, p. 2387-2398, Vol. 26, No. 6
0022-538X/06/$08.00+0     doi:10.1128/MCB.26.6.2387-2398.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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