NDLI: C-terminal cleavage of DeltaNp63alpha is associated with TSA-induced apoptosis in immortalized corneal epithelial cells.

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C-terminal cleavage of DeltaNp63alpha is associated with TSA-induced apoptosis in immortalized corneal epithelial cells.

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C-terminal cleavage of DeltaNp63alpha is associated with TSA-induced apoptosis in immortalized corneal epithelial cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Robertson, Danielle M. Ho, Su-Inn Cavanagh, H. Dwight
Description	Country affiliation: United States Author Affiliation: Robertson DM ( Department of Ophthalmology, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-9057, USA. danielle.robertson@utsouthwestern.edu)
Abstract	PURPOSE: In the central human corneal epithelium, loss of DeltaNp63 occurs in all surface epithelial cells preparing to undergo desquamation, suggesting a potential role for DeltaNp63 isoforms in mediating surface cell apoptotic shedding. In this study, the authors investigated a role for DeltaNp63 isoforms in caspase-mediated apoptosis in a telomerase-immortalized corneal epithelial cell line. METHODS: For in vitro studies, hTCEpi cells were cultured in KGM-2 serum-free culture media containing 0.15 mM calcium. To assess dynamic protein interactions among individual DeltaNp63 isoforms, DeltaNp63-EGFP expression plasmids were transiently expressed in hTCEpi cells and evaluated by FRAP. Trichostatin-A (TSA; 3.31 muM) was used to induce cell death as measured by caspase activity. Cleavage and loss of endogenous DeltaNp63alpha, DeltaNp63-EGFP expression plasmids, and p53 were assessed after treatment with TSA and siRNA. RESULTS: Transient expression of DeltaNp63-EGFP alpha and beta isoforms resulted in the formation of a smaller isoform similar in size to DeltaNp63gamma-EGFP. FRAP demonstrated that DeltaNp63alpha-EGFP has greater immobile fraction than beta or gamma. TSA induced caspase-mediated apoptotic pathways; caspase induction was accompanied by a decrease in endogenous DeltaNp63alpha and p53. TSA upregulated DeltaNp63-EGFP plasmid expression; this was accompanied by a selective increase in cleavage of DeltaNp63alpha-EGFP. siRNA knockdown of DeltaNp63alpha correlated with a reduction in p53 independently of TSA. CONCLUSIONS: DeltaNp63alpha is the dominant active isoform in corneal epithelial cell nuclei. Loss of DeltaNp63alpha occurs during apoptotic signaling by cleavage at the C terminus. The corresponding loss of p53 suggests that a significant relationship appears to exist between these two regulatory proteins.
ISSN	01460404
e-ISSN	15525783
DOI	10.1167/iovs.09-4919
Journal	Investigative Opthalmology & Visual Science
Issue Number	8
Volume Number	51
Language	English
Publisher	Association for Research in Vision and Ophthalmology
Publisher Date	2010-08-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Apoptosis Drug Effects Epithelium, Corneal Pathology Hydroxamic Acids Pharmacology Protein Synthesis Inhibitors Trans-activators Physiology Tumor Suppressor Proteins Blotting, Western Caspase 3 Metabolism Cell Culture Techniques Fluorescence Recovery After Photobleaching Gene Silencing Green Fluorescent Proteins Genetics Plasmids Protein Isoforms Rna, Small Interfering Recombinant Fusion Proteins Transcription Factors Transfection Tumor Suppressor Protein P53 Up-regulation Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Discipline Ophthalmology
Content Type	Text
Resource Type	Article
Subject	Ophthalmology Sensory Systems Cellular and Molecular Neuroscience

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