December 2002
Volume 43, Issue 13
ARVO Annual Meeting Abstract  |   December 2002
Two Sequences in the 3'UTR of ß-PDE Control Gene Expression in Human Cells and Xenopus Laevis
Author Affiliations & Notes
  • MR Verardo
    UCLA Neuroscience IDP and Jules Stein Eye Institute Los Angeles CA
  • NI Piriev
    Jules Stein Eye Institute UCLA Los Angeles CA
  • BE Knox
    SUNY HES Syracuse NY
  • DB Farber
    Jules Stein Eye Institute UCLA Los Angeles CA
  • Footnotes
    Commercial Relationships   M.R. Verardo, None; N.I. Piriev, None; B.E. Knox, None; D.B. Farber, None. Grant Identification: EY02651
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 3654. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      MR Verardo, NI Piriev, BE Knox, DB Farber; Two Sequences in the 3'UTR of ß-PDE Control Gene Expression in Human Cells and Xenopus Laevis . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3654.

      Download citation file:

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

  • Supplements

Abstract: : Purpose: Previously, we reported that in Y-79 retinoblastoma cells two 100 nt fragments from the 3'untranslated region (3'UTR) of rod cGMP-phosphodiesterase ß-subunit (ß-PDE) enhance luciferase reporter gene expression above that produced by the complete 3'UTRs of ß-PDE, SV-40, and other photoreceptor specific genes. The purpose of our current work was to determine if this effect was specific to Y-79 cells and the SV-40 promoter that was used to drive luciferase expression of the constructs. Further, we wanted to find out if the effect of these 3'UTR fragments on gene expression was conserved across species. Methods: The two 100 nt fragments from the 3'UTR of ß-PDE were cloned downstream of the luciferase gene in a modified pGL3 vector under control of the ß-PDE promoter. These constructs were transfected ex-vivo in Xenopus laevis tadpole heads and also in Y-79 cells. HEK 293, HeLa, and SY5Y cells were transfected with the SV-40 promoter constructs. Luciferase assays were performed on harvested cell and tissue lysates. Results: With constructs driven by the SV-40 promoter, the entire 3'UTR of ß-PDE decreased luciferase expression in all cell lines tested, but both 100 nt fragments increased it in HEK293 and had no effect in HeLa cells. Only one 3'UTR fragment increased luciferase expression in SY5Y cells. Interestingly, constructs driven by the ß-PDE promoter transfected ex-vivo in Xenopus heads reproduced the observations obtained in vitro with Y-79 retinoblastoma cells. Conclusion: Although the complete 3'UTR of ß-PDE decreased luciferase expression, the two 100 nt fragments increased it differently, depending on the cell lines used for transfection. This implies cell specific mechanisms may control expression by elements of the ß-PDE 3'UTR. Our results also show that the effect of the 3'UTR fragments is conserved from human to amphibians, suggesting that these sequences may have an important role in gene regulation in vertebrate retinas.

Keywords: 417 gene/expression • 517 photoreceptors • 476 molecular biology 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.