developmental genetics
DESCRIPTION
Developmental Genetics. Overview of gene structure Mechanisms of gene regulation Drosophila: Genetics model for Developmental Biology. What is a gene?. -10. -35. +1. AAAAAA. UAA. AUG. Regulatory Promoter. 3 UTR. Core Promoter. CDS. Developmental Genetics. - PowerPoint PPT PresentationTRANSCRIPT
Developmental Genetics Overview of gene structure Mechanisms of gene regulation Drosophila: Genetics model for Developmental
Biology
What is a gene?
CDSRegulatoryPromoter
Core Promote
r
-10-35 +1
3 UTR
AAAAAAAUG UAA
Developmental Genetics Differentiation of
cells requires regulation of specific genes.
Timing of Gene Regulation can determine when a tissue develops or when cells undergo Apoptosis.
How can we regulate gene expression?
Chromatin Remodeling
Histone Code: Modification of Histone tails. Acetylation, methylation, and
phosphorylation. Charged residues in tail.
Histone Methylation Methylation of residues on
H3 tail
Methylation of H3K4(meH3K4) + high acetylation of H3/H4 tails)= active transcription.
meH3K9+low acetylation of H3/H4= highly repressed
meH3K9, meH3K27, meH4K20 =highly repressed.
Example: Hox genes Hox genes are
responsible for determining the fate of segments along the anterior-posterior axis.
Trimethylation of H3K27 represses Hox genes.
Demethylase for me3H3K27 is recruited in differentiated cells to open up the promoters for Hox genes.
Epigenetic Memory Trithorax- keeps genes active.
Counteract Polycombs Modify nucleosomes Keep H3K4 trimethylated
Polycomb- keeps genes inactivated. H3K27 methylase Bind meH3 tails
Methylation of DNA Once gene is turned on and needs to stay on CpG-usually associated with repression.
b-globin promoter. Methylation status of promoter can change
during development. Ex. Embryonic hemoglobin (e-globin) fetal
hemoglobin (g-globin)
Methylation and Transcriptional repression Can block
transcription factor binding
Recruit Histone methylases and acetylases Ex. MeCP2
Dnmt3 and Dnmt1 result in the heritability of methylated DNA regions in progeny cells.
Methylation can inactivate entire chromosomes.
Transcriptional regulation cis factors trans factors
Promoters/enhancers Core promoter
RNA Pol II binds to this region. Directly upstream of transcription start site.
Regulatory Promoter Region upstream of core promoter Contains enhancers/regulatory binding sites Highly modular.
Promoter fusions Identifying new
enhancers Reporter genes: b-gal
and GFP
Enhancer Modulation Multiple transcription
factors can bind to a single Promoter region.
Pax6 enhancer region-differential transcription factor binding for different tissue types.
Pax61 Core promoter1 2 3 4
Pay it forward Pax6 is a
transcriptional regulator.
Pax6 works with Sox2 and L-Maf to regulate the crystallin gene.
Pax6 also regulates Pancreas related genes Insulin, glucagon, and
somatostatin Pax6 regulates itself!
Transcription Factors Core Promoter
TFIID/TBP stabilized by TFIIA
TFIIH binds, RNA Pol II with TFIIE/TFIIF binds subsequently.
TFIIH phosphroylates RNA Pol II CTD.
Regulatory Promoter Wide array of factors
bind to enhancer regions. (Pax6 regulation).
Transcription Factor Domains. DNA binding domain Transactivating
Domain Protein-Protein
Interaction domain.
MITF TranscriptionFactor- Ear developmentAnd Pigment production
“Pioneer” transcription factors. Pioneer transcription
factors can penetrate repressed chromatin.
FoxA1 Pax7 Pbx
Gene Silencing Neural restrictive silencer
element (NRSE) found represses neural genes in non-neural cells. NRSF binds to this element.
L1 is critical for brain development.
Transgene reporter with NRSE= neural tissue expression only
Transgene reporter without NRSE= all tissues.
Regulation of mRNA mRNA must undergo splicing for protein to be
made. Regulation of mRNA
Determination of which transcripts get translated Splicing combinations to make new protein.
Splicing: different gene products from one gene. Bcl-x transcript.
Bcl-xL and Bcl-xS
Tropomysin Leads to multiple
proteins that differ in function and location.
Splicing enhancers-found in specific tissues.
Splicing can silence as well- splice excludes an exon from mRNA.
Translational Regulation Stored oocyte mRNAs-
selective inhibition of translation.
Genes turned on quickly post-fertilization.
Relies on the recruitment of proteins involved in making a polyA tail.
Drosophila Bicoid gene product.
miRNA miRNA ancient form
of regulation. Protection against
viral pathogens Quick regulation and
turnover of mRNA.
Drosophila Development: Genetics at work!
Development of Drosophila
Anterior-Posterior Polarity Starts during oogenesis Gurken mRNA
transferred to oocyte. Signal sent back to
oocyte post Gurken/Torpedo interaction.
Par-1 mediated organization of microtubules to posterior side of oocyte. Minus end (posterior) and plus end (anterior).
Anterior-Posterior Polarity
Dorsal-Ventral Patterning
Dorsal Gradient