May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Pax–6 and Sox11 as Possible Regulators of the Optimedin Gene Activity
Author Affiliations & Notes
  • O.V. Grinchuk
    Lmdb, NEI NIH, Bethesda, MD
  • X. Wu
    Lmdb, NEI NIH, Bethesda, MD
  • K. Zbinek
    Lmdb, NEI NIH, Bethesda, MD
  • S.I. Tomarev
    Lmdb, NEI NIH, Bethesda, MD
  • Footnotes
    Commercial Relationships  O.V. Grinchuk, None; X. Wu, None; K. Zbinek, None; S.I. Tomarev, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 653. doi:
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      O.V. Grinchuk, X. Wu, K. Zbinek, S.I. Tomarev; Pax–6 and Sox11 as Possible Regulators of the Optimedin Gene Activity . Invest. Ophthalmol. Vis. Sci. 2004;45(13):653.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Optimedin is an olfactomedin–containing protein that may interact with myocilin. Optimedin gene is expressed in the retina, tissues of the eye angle and brain. We investigated possible involvement of several transcription factors in the regulation of optimedin promoter activity. Methods: Promoter fragments of the mouse optimedin gene were cloned into the pGL3 vector and their activity was tested in vitro after transfection into COS7 cells using luciferase as a reporter. Interaction of Pax–6 with a putative binding site in the proximal optimedin gene promoter was studied by gel shift assays. Mutations in a putative Pax–6 binding site were introduced by site–directed mutagenesis. Results: –2829/+98 and –136/+24 optimedin promoter fragments showed 6 fold stimulation over the promoterless vector after transfection into COS7 cells. Pax6 stimulated luciferase activity driven by these promoter fragments 8–10 fold. A putative Pax–6 binding site was identified in the optimedin promoter at position –86/–70. Pax–6 is able to bind to this site as judged by gel shift assay. A mutation GCG→ttc(position –76 /–73) in this site completely eliminated Pax–6 binding. Similar mutation did not change the basal activity of the optimedin promoter fragments in COS–7 cells. However, Pax–6 stimulated the mutated promoter two fold only. Co–transfection experiments with several transcription factors expressed in the retina and brain demonstrated that Brn3b, Math5, NeuroD and Sox2 did not show any stimulatory effect on the optimedin promoter fragments while Sox11 stimulated the activity of –2829/+98 and –136/+24 promoter fragments 19 and 12 fold, respectively. Putative Sox–binding sites were identified in the optimedin proximal promoter. Experiments are in progress to test binding of Sox11 to these sites. Conclusions: Our data suggest that Pax–6 and Sox11 may be involved in the regulation of the optimedin promoter in the retina and brain in vivo.

Keywords: ganglion cells • retina • transcription factors 
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