Talk1 Drosophila Talk 205 L
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Transcript of Talk1 Drosophila Talk 205 L
Characterizing eve(PJ4)
Biology 205L: Experiments in Developmental Biology Fall 2006
Myung Ko, Jasmine Lau, Joshua Mendoza-Elias
Gene Regulatory Network
Insert Figure 9.26 (Gilberts)
eve
wg
runt odd
en
hh
TF
+
Consequences of wg expression
Jones & Bejsovec (2003)
An anomaly: eve(PJ4)wg(NE2)
• Gastrulation- wingless protein expression
• Cuticle patterning- wingless expression
Questions:
• Is eve(PJ4) an amorph? • Role of wg(NE2) in double mutant?
Hypothesis:
• eve(PJ4) is an amorph• wg(NE2) causes wg expression to persist
Materials and Methods
Statistical Methods: 2-test, t-test
Wild-type
eve(3)
eve(PJ4) eve(PJ4)wg(NE2)
AB-staining: anti-wg antibody Stages: 9, 10,
11
Cuticle Preparations
Part I: Is eve(PJ4) an amorph?
Are we seeing the right ratio?
• eve (PJ4) • eve(3)
WT Mutant
Ratio
3 1
n 17 8
WT Mutant
Ratio
3 1
n 45 162 value = 0.333 p < 0.05
2 value= 0.022 p< 0.01
Stage 9eve(PJ4)
Mean: 5.89
(n=9)
eve(3)
Mean:6.21
(n=9)
WTWT
WT
Mean: 13.2
(n= 5)
P= 0.31
P= 0.00
P= 0.00
Stage 10
eve (PJ4)
Mean: 5.86
(n=7)
eve(3)
Mean: 5.5
(n=12)
WT
Mean:13.5
(n= 17)
P= 0.47
P= 0.00
P= 0.00
Stage 11
eve(PJ4)
Mean: 5.3
(n=3)
eve(3)
Mean: 5
(n=3)
WT
Mean: 14.3
(n=3)
P= 0.65
P= 0.00
P= 0.00
Mean: 5.89
Mean:6.21
Mean: 5.86
Mean: 5.5
Mean: 5.3
Mean: 5
eve(PJ4)
Stage 9 Stage 10 Stage 11
eve (3)
P= 0.31 P= 0.47 P= 0.65
Cuticles
eve(PJ4) eve(3)
Summary
• eve(PJ4) behaves like an amorph
• As embryos mature,
- decreasing # of stripes
- stripes are less defined
- increasing similarity b/w eve(PJ4) and eve(3)
Part II: Role of wg(NE2) in eve(PJ4)wg(NE2)
wg(NE2) to the rescue??
Are we seeing the right ratio?
• eve(PJ4) • eve(PJ4)wg(NE2)
WT Mutant
Ratio
3 1
n 17 8
WT +/- Mutant
Ratio
1 2 1
n 8 18 10
2 value = 0.33 p < 0.05
2 value= 0.89 p < 0.01
Stage 9
eve(PJ4)
Mean: 5.89
(n=9)
eve(PJ4)wg(NE2)
Mean:5.86
(n=7)
WT
Mean: 13.2
(n= 5)
P= 0.95
P= 0.00
P= 0.00
Stage 10
eve(PJ4)
Mean: 5.86
(n= 7)
eve(PJ4)wg(NE2)
Mean:5.7
(n= 10)
WT
Mean: 13.5
(n= 17)
P= 0.76
P= 0.00
P= 0.00
Stage 11eve(PJ4)
Mean: 5.3
(n=3)
eve(PJ4)wg(NE2)
Mean: 6.14
(n= 7)
WT
Mean: 14.3
(n= 3)
P= 0.12
P= 0.00
P= 0.00
Mean: 5.89
Mean:5.86
Mean: 5.86
Mean:5.7
Mean: 5.3
Mean: 6.14
P= 0.95 P= 0.76 P= 0.12
eve(PJ4)
Stage 9 Stage 10 Stage 11
eve(PJ4) wg(NE2)
Cuticles
eve(PJ4) eve(PJ4)wg (NE2)
Summary
• Effects of wg(NE2)
- slower decrease in wg expression
- more defined stripes
- increasing deviation of eve(PJ4)wg(NE2) from eve(PJ4)
- double mutant still behaves like an amorph
Discussion
• EVX1, a human homeobox gene homologous to eve
- Transcriptional repressor: specifies A-P axis in embryos
- EVX1 -/- : increased oncogenic potential
Future Directions
• 0-4 hours Ab staining• 0-4 hours and 4-8 hours In Situ
hybridization• eve(1) mutants• Stages 12 to 15 (cuticle secretion)• Sequence eve(PJ4)
Acknowledgements
• Dr. Bejsovec
• Dr. Perz-Edwards
• Wendy Beane
• Monica Zhang
• Our wonderful classmates!
Thank you!
References
1. Paul M. Mcdonald, Phil Ingham and Gary Struhi. Isolation, Structure, and Expression of even-skipped: A Second Pair-Rule Gene of Drosophila Containing a Homeo Box. Cell: Vol. 47, 721-734 (1986)
2. Marcel van den Heuvel, Roel Nusse, Paul Johnston, and Peter A. Lawrence. Distribution of the wingless Gene Product in Drosophila Embryos: A Protein Involved in Cell-Cell Communication. Cell: Vol. 59, 739-749 (1989)
3. Herman A. Dierick and Amy Bejsovec. Functional analysis of Wingless reveals a link between intercellular ligand transport and dorsal-cell-specific signaling. Development: 125, 4729-4738 (1998)
4. Whitney M Jones and Amy Bejsovec. Wingless Signaling: An Axin to Grind. Current Biology: Vol.13, R479-R481 (2003)
5. Antonio Faiella, Maurizio D'Esposito, Marco Rambaldi, Dario Acampora, Silvia Balsofiore, Anna Stornaiuolo, Antonio Mallamaci, Enrica Migliaccio, Massimo Gulisano, Antonio Simeone and Edoardo Boncinelli. Isolation and mapping of EVX1, a human homeobox gene homologous to even-skipped, localized at the 5' end ofHOXi locus on chromosome 7. Nucleic Acids Research: Vol. 19, No. 23 6541-6545 (1991).
6. Chi Li and James Manley. Even-skipped Represses Transcription by Binding TATA Binding Protein and Blocking the TFIID-TATA Box Interaction. Molecular and Cellular Biology: Vol. 18, No.7, p. 3771–3781 (1998).
7. Paola Briata, Rinke Van De Werken, Irma Airoldi, Cristina Ilengo, Erica Di Blas, Edoardo Boncinelli, and Giorgio Corte. Transcriptional Repression by the Human Homeobox Protein EVX1 in Transfected Mammalian Cells. The Journal of Biological Chemistry: Vol. 270, No. 46, pp. 27695–27701, (1995).