Pax-6 interactions with TATA-box-binding protein and retinoblastoma protein

Ales Cvekl, Fatah Kashanchi, John N. Brady, Joram Piatigorsky

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

PURPOSE. To identify proteins that physically interact with Pax-6, a paired domain- and homeodomain (HD)-containing transcription factor that is a key regulator of eye development. METHODS. Protein-protein interactions involving Pax-6, TATA-box-binding protein (TPB), and retinoblastoma protein were studied using affinity chromatography with Pax-6 as ligand, glutathione- S-transferase (GST) pull-down assays, and immunoprecipitations. RESULTS. The authors have shown that Pax-6 is a sequence-specific activator of many crystallin genes, all containing a TATA box, in the lens. Others have shown that lens fiber cell differentiation, characterized by temporally and spatially regulated crystallin gene expression, depends on retinoblastoma protein, in the present study it was shown that Pax-6 interacted with the TBP, the DNA-binding subunit of general transcription complex TFIID. GST pull-down assays indicated that this interaction was mediated by the Pax-6 HD, with a substantial role for its N-terminal arm and first two α-helices. The experiments also indicated a binding role for the C-terminal-activation domain of the protein. In addition, the present study showed that the HD of Pax-6 interacted with retinoblastoma protein. Immunoprecipitation experiments confirmed retinoblastoma protein/Pax-6 complexes in lens nuclear extracts. CONCLUSIONS. Blending the present results with those in the literature suggests that Pax-6 and retinoblastoma protein participate in overlapping regulatory pathways controlling epithelial cell division, fiber cell elongation, and crystallin gene expression during lens development.

Original languageEnglish (US)
Pages (from-to)1343-1350
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume40
Issue number7
StatePublished - 1999

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TATA-Box Binding Protein
Retinoblastoma Protein
Crystallins
Lenses
Glutathione Transferase
Immunoprecipitation
Transcription Factor TFIID
Gene Expression
TATA Box
Proteins
Affinity Chromatography
Cell Division
Cell Differentiation
Arm
Transcription Factors
Epithelial Cells
Ligands
DNA
Genes

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Pax-6 interactions with TATA-box-binding protein and retinoblastoma protein. / Cvekl, Ales; Kashanchi, Fatah; Brady, John N.; Piatigorsky, Joram.

In: Investigative Ophthalmology and Visual Science, Vol. 40, No. 7, 1999, p. 1343-1350.

Research output: Contribution to journalArticle

Cvekl, Ales ; Kashanchi, Fatah ; Brady, John N. ; Piatigorsky, Joram. / Pax-6 interactions with TATA-box-binding protein and retinoblastoma protein. In: Investigative Ophthalmology and Visual Science. 1999 ; Vol. 40, No. 7. pp. 1343-1350.
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