Facts:

-Bacteria (4000 genes) 90% genes are involved with growth and prolifera=on 10% genes dieren=a=on have genes organized in operons promoters are small and very specic -Yeasts (4500 genes) 90% of genes are involved with growth and prolifera=on 10% of genes dieren=a=on have genes dispersed promoters are medium size and very specic -Mul=cellular (25000 genes) 20% genes involved with growth and prolifera=on 40% genes involves with development 40% genes involved with specialized func=ons

How is a mul=cellular organism constructed ?

Highly specialized cells

To=potent cell

zygote

Do cells loose genes as they are undergoing development and dieren=a=on ?

Cloning says NO !!

Expression of dierent genes in dierent cells types

DNA micro-arrays

Dierent proteins are produced in dierent =ssues at dierent =mes

2D-SDS protein gel electrophoresis

Regula=on of gene expression can occur at many dierent stages

Three key components in the regula=on of gene expression: 1.

Signals: hormones (estrogen, testosterone, progesterone) Growth factors (NGF, CSF, Interferon,) environment (drugs, toxic substances, metals)

2. Level of response transcrip=on mRNA processing mRNA transport and localiza=on mRNA transla=on mRNA stability Protein ac=vity 3. Mechanisms Transac=va=ng proteins Methyla=on X-inac=va=on

Transcrip=on is ini=ated by proteins recognizing the Promoter of a gene

 Transcrip=on factors bind specic regulatory cis-ac=ng sequences and promote transcrip=on RNA poly II -5000pb +1 TATA +1 TATA mRNA

OFF

-5000pb

mRNA

low

-5000pb

+1 TATA

mRNA

ON

Enhancer

Basic promoter

Transcrip=on factors

Transcrip=on factors recognise short DNA sequences

 Families of DNA-binding transcrip=on factors

- DNA binding proteins

 Dierent types of regula=on

 Organiza=on of a eukaryo=c promoter

 Organiza=on of a eukaryo=c gene regulatory region Basic Promoter Enhancer 5 -5000pb TATA Gene Enhancer 3

Enhancers have variable: -distance -posi=on -orienta=on

Basic promoter has xed: - posi=on - distance - orienta=on

 Ac=vity of enhancers at a distance

 Ac=vity of enhancers at a distance

 Sequence of events during ac=va=on of transcrip=on

Histone acetyla=on gene transcrip=on

 Assembly of transcrip=onal complexes at the promoter

 Possible post-transla=onal modica=ons of transcrip=on factors

 Dieren=al expression of fetal and maternal globin genes

G A

Low Oxigen pressure, high anity High Oxigen pressue, lower anity

Transla=onal regula=on during embryogenesis

 Glucoccor=coids diuse to the nucleus and could work at many dierent promoters

 Alterna=ve splicing: how to get more proteins out of your genome 5 DNA Nuclear Transcripts mRNAs Proteins 1 1 1 2 2 2 3 3 3 4 calcitonin 4 calcitonin 4 calcitonin 4 calcitonin
AAAA

5 CGRP

5 CGRP

1 2 3 Thyroid

1 2 3 Neurons

5 CGRP

AAAA

30,000 human genes probably give more than 100,000 dierent proteins

Alterna=ve promoters and splicing a-amylase

Regula=ng stability of mRNA

Promoter dele=on studies

Promoter dele=on studies with GFP in Whole animals Conec=ve =ssue growth factor

Cellular Stress Response

Many genes upregulated

Control of GAL regula=on in yeast

- Ac=vator protein GAL4 - Inhibitory protein GAL80

 Eukaryo=c promoters are very complex and receive mul=ple signals

Complexity of eukaryo=c promoters

Nuclear matrix associa=on of promoters

Genoma wide mapping SARs/MARs

 Nature of insulators

 The eect of cytosine methyla=on upon gene expression

Heritable pakerns of expression maintained though DNA replica=on

 The eect of cytosine methyla=on upon gene expression

 Housekeeping genes

CpG repe==ve blocks usually mark genes cons=tu=vely expressed

Drosophila Developmental Gene=cs

 Combinatorial gene control during development

 Combinatorial gene control during development

 The epigene=c landscape

Conrad Waddington (1905-1975)

 The epigene=c landscape

Conrad Waddington (1905-1975)

The epigene=c landscape

Epigene=c gene regula=on