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Understanding Gene Expression in Developmental Biology

Explore the cellular functions crucial for development, gene expression steps, and the regulatory mechanisms of transcription and protein turnover. Learn about the role of transcription factors, histone modifications, and maintaining gene expression states. Discover how genes encode developmental programs and why cells behave differently. Techniques like Southern blotting and in situ hybridization are discussed for monitoring gene expression in specific cells.

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Understanding Gene Expression in Developmental Biology

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  1. March 3, 2010 Introduction to development Gene expression

  2. What cellular functions are needed to carry out development?

  3. What cellular functions are needed to carry out development? Divide Grow Differentiate Die Move Adhere Secrete Signal Cell biological properties need to be coordinated in space and time

  4. Where is the “program” for development encoded?

  5. Where is the “program” for development encoded? In the genes

  6. Why do different cells behave differently?

  7. Why do different cells behave differently? Have different genes? Have different histories? Experience different environments? Chance? Have different gene expression states?

  8. Why do different cells behave differently? Have different genes? Have different histories? Experience different environments? Chance? Have different gene expression states?

  9. 05_02_DNA.jpg

  10. 05_10_Genes_info.jpg Genes Carry out functions

  11. Steps in gene expression 07_37_Protein.produc.jpg

  12. Regulation of gene expression 08_03_control.steps.jpg mRNA localization control (tether mRNA to localized proteins) mRNA turnover control (microRNAs) Protein turnover control (ubiquitylation and proteasome digestion) Protein localization control

  13. Regulation of gene expression 08_03_control.steps.jpg mRNA localization control (tether mRNA to localized proteins) mRNA turnover control (microRNAs) Protein turnover control (ubiquitylation and proteasome digestion) Protein localization control

  14. Regulation of transcription: Parts of a gene coding strand

  15. Regulation of transcription 08_13_gene.activation.jpg

  16. Transcription factors

  17. 05_24_Chromatin pack.jpg Nucleosomes (histones) package DNA

  18. Histone modifications affect gene expression 05_30_histone tails.jpg Can affect recruitment of transcription factors to promoter

  19. Some transcription factors affect histones 08_14_chromatin.struc.jpg

  20. Multiple transcription factors regulate most genes 08_15_Reg. proteins.jpg

  21. Modularity of the Drosophila even-skipped promoter 08_18_reporter.gene.jpg

  22. Coordinated regulation of multiple genes Developmental functions

  23. Signals can regulate activity of transcription factors

  24. Maintaining gene expression states 08_23_cell.memory.jpg (a positive feedback loop)

  25. Maintaining gene expression states 08_24_chromatin.state.jpg

  26. The histone code also needs to be maintained in daughter cells

  27. Maintaining gene expression states – DNA methylation

  28. How does one monitor which genes a particular cell expresses?

  29. Southern blot 10_14_1_Southrn.blotting.jpg

  30. 10_14_2_Southrn.blotting.jpg Southern blot – DNA on blot Northern blot – RNA on blot

  31. Northern blot hybridization (tests one gene at a time)

  32. In situ hybridization of developing flowers with ARF6 probe

  33. Rb-Related expression in Arabidopsis embryos by in situ hybridization From Wildwater et al., Cell 123: 1337-1349 (2005)

  34. 08_18_reporter.gene.jpg Promoter:reporter fusion gene in a transgenic fly embryo

  35. PARF6::ARF6::GUS fusion expression in flowers and ovules Promoter:protein:reporter fusion gene – reveals protein location

  36. Immunolocalization of Snail and Twist proteins in Drosophila embryos anti-Snail anti-Twist Kosman et al., Science 254: 118-122 (1991)

  37. Utility of looking at expression of single genes at a time: Markers of cell type, differentiation Visualization of regulatory events Utility of looking at expression of many genes at once: Global view of tissue identity Comparing different tissues or states Global view of regulatory events

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