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Alternative Splicing in the TCF/LEF
Family of Transcription Factors
Elizabeth Kessell - Professor Scott Roy

San Francisco State University - NSF REU Program
The TCF/LEF Family

 •Transcription factors

 •Part of Wnt pathway

 •4 family members - TCF7, TCF7-l1, TCF7-l2, LEF1

 •Transcripts expressed differently in different tissues/developmental stages




            Image Credits: Wikipedia Commons - http://en.wikipedia.org/wiki/File:Protein_TCF7L2_PDB_1jdh.png , my own work
Dominant Negative Isoforms


•Without a B-catenin binding
site, the pathway remains off

•This serves as a regulatory
mechanism

•Proteins lacking a binding site
are created through alternative
splicing
Introns, Exons, and Alternative Splicing
An Evolutionary Question




     How far back do the dominant negative
                   forms go?

             How did they evolve?
What we would expect to see
Methods



•Using sequence mapping to match intron sequence to cDNA data
•Looking for a mature RNA that contains sequence previously thought to be intron
•Data from danio rerio (fish), xenopus tropicalus (frog), anolis carolinensis (lizard)
Results




   None of the data tested showed an alternative 1st exon
               (for any of the species tested)
Interpretation

 •Strong negative result

 •Implies that TCF7l2 and LEF1 independently
 evolved dominant negative forms in mammals

 •Inconclusive -- more data?
Moving Forward

 •Get more cDNA data

 •Learn more about the negative isoforms in mammals

 •Look beyond vertebrates
Acknowledgements

   Many thanks to Professor Roy for mentorship and advice, as
   well as to San Francisco State University and the

   National Science Foundation for funding and
   support.
References

•Weise, et al, Alternative splicing of Tcf7l2 transcripts generates protein variants with differential promoter-binding and
transcriptional activation properties at Wnt/!-catenin targets, Nucleic Acids Research, 2010

•Huber, et al, Nuclear localization of !-catenin by interaction with transcription factor LEF-1, Mechanisms of
Development 1996

•Lin, et al, Expression of the AmphiTcf Gene in Amphioxus: Insights Into the Evolution of the TCF/LEF Gene Family
During Vertebrate Evolution, Developmental Dynamics, 2006

•Klingel, et al, Subfunctionalization and neofunctionalization of vertebrate Lef/Tcf transcription factors, Developmental
Biology, 2012

•Hovanes, et al, The human LEF-1 gene contains a promoter preferentially active in lymphocytes and encodes multiple
isoforms derived from alternative splicing, Nucleic acids research, 2000

•Vacik and Lemke, Dominant-negative isoforms of Tcf/Lef proteins in development and disease, Cell Cycle, 2011
Alternative Splicing in the TCF/LEF
Family of Transcription Factors
Elizabeth Kessell - Professor Scott Roy

San Francisco State University - NSF REU Program

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TCF/LEF talk

  • 1. Alternative Splicing in the TCF/LEF Family of Transcription Factors Elizabeth Kessell - Professor Scott Roy San Francisco State University - NSF REU Program
  • 2. The TCF/LEF Family •Transcription factors •Part of Wnt pathway •4 family members - TCF7, TCF7-l1, TCF7-l2, LEF1 •Transcripts expressed differently in different tissues/developmental stages Image Credits: Wikipedia Commons - http://en.wikipedia.org/wiki/File:Protein_TCF7L2_PDB_1jdh.png , my own work
  • 3. Dominant Negative Isoforms •Without a B-catenin binding site, the pathway remains off •This serves as a regulatory mechanism •Proteins lacking a binding site are created through alternative splicing
  • 4. Introns, Exons, and Alternative Splicing
  • 5. An Evolutionary Question How far back do the dominant negative forms go? How did they evolve?
  • 6. What we would expect to see
  • 7. Methods •Using sequence mapping to match intron sequence to cDNA data •Looking for a mature RNA that contains sequence previously thought to be intron •Data from danio rerio (fish), xenopus tropicalus (frog), anolis carolinensis (lizard)
  • 8. Results None of the data tested showed an alternative 1st exon (for any of the species tested)
  • 9. Interpretation •Strong negative result •Implies that TCF7l2 and LEF1 independently evolved dominant negative forms in mammals •Inconclusive -- more data?
  • 10. Moving Forward •Get more cDNA data •Learn more about the negative isoforms in mammals •Look beyond vertebrates
  • 11. Acknowledgements Many thanks to Professor Roy for mentorship and advice, as well as to San Francisco State University and the National Science Foundation for funding and support.
  • 12. References •Weise, et al, Alternative splicing of Tcf7l2 transcripts generates protein variants with differential promoter-binding and transcriptional activation properties at Wnt/!-catenin targets, Nucleic Acids Research, 2010 •Huber, et al, Nuclear localization of !-catenin by interaction with transcription factor LEF-1, Mechanisms of Development 1996 •Lin, et al, Expression of the AmphiTcf Gene in Amphioxus: Insights Into the Evolution of the TCF/LEF Gene Family During Vertebrate Evolution, Developmental Dynamics, 2006 •Klingel, et al, Subfunctionalization and neofunctionalization of vertebrate Lef/Tcf transcription factors, Developmental Biology, 2012 •Hovanes, et al, The human LEF-1 gene contains a promoter preferentially active in lymphocytes and encodes multiple isoforms derived from alternative splicing, Nucleic acids research, 2000 •Vacik and Lemke, Dominant-negative isoforms of Tcf/Lef proteins in development and disease, Cell Cycle, 2011
  • 13. Alternative Splicing in the TCF/LEF Family of Transcription Factors Elizabeth Kessell - Professor Scott Roy San Francisco State University - NSF REU Program

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