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• Control of Gene Expression
Chip array analysis tumors
1800 genes
Red- up Black- same Green down
Blue is different
Gene expression is regulated at many levels
How was this worked out ? Bacteria RNA polymerase + a factor Eucaryotes Didn’t know what a promotor was , Ad major late . Extract that works ,looking for sigma factor Manley Roeder. Reinberg, Tijan
DNA
A Positive Regulatory Element Maps Between -115 and -153 -1004
Luc
-890
Luc
-681
Luc
-428
Luc
-248
Luc
-228
Luc
-195 -153 -115
Luc Luc Luc 0
10
20
30
40
50
Fold Activation
60
How do gene regulatory proteins work? What is seen on the DNA ? Sequence specific but do not unwind
Most proteins bind to the major groove
What are structural characteristics of the gene regulatory proteins ?
The simplest and most typical is the helix-turn-helix
The homeodomain is a helix-turn -helix protein
The zinc finger - reiterations can strengthen interaction
The glucocorticoid receptor has two zinc fingers
A specific regulatory factor for each gene ? NO!! Heterodimers-leucine zippers
The Heliz-loop-helix motif mediates Dimerization and DNA binding Another solution to number problem
Complex expression patterns are a result of the concerted action of discrete simpler elements
Regulation of transcription by covalent modification of DNA
Methylation CpG May explain heritable patterns of gene expression
Regulation of gene expression by RNA itself Do transcription factors contain RNA ?
Regulation of processing and expression by RNA
Chip array analysis tumors
1800 genes
Red- up Black- same Green down
Blue is different
Gene expression is regulated at many levels
How was this worked out ? Bacteria RNA polymerase + a factor Eucaryotes Didn’t know what a promotor was , Ad major late . Extract that works ,looking for sigma factor Manley Roeder. Reinberg, Tijan
DNA
A Positive Regulatory Element Maps Between -115 and -153 -1004
Luc
-890
Luc
-681
Luc
-428
Luc
-248
Luc
-228
Luc
-195 -153 -115
Luc Luc Luc 0
10
20
30
40
50
Fold Activation
60
How do gene regulatory proteins work? What is seen on the DNA ? Sequence specific but do not unwind
Most proteins bind to the major groove
What are structural characteristics of the gene regulatory proteins ?
The simplest and most typical is the helix-turn-helix
The homeodomain is a helix-turn -helix protein
The zinc finger - reiterations can strengthen interaction
The glucocorticoid receptor has two zinc fingers
A specific regulatory factor for each gene ? NO!! Heterodimers-leucine zippers
The Heliz-loop-helix motif mediates Dimerization and DNA binding Another solution to number problem
Complex expression patterns are a result of the concerted action of discrete simpler elements
Regulation of transcription by covalent modification of DNA
Methylation CpG May explain heritable patterns of gene expression
Regulation of gene expression by RNA itself Do transcription factors contain RNA ?
Regulation of processing and expression by RNA
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