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10/25/2014 1
To know and explain: 
• Regulation of Bacterial Gene Expression 
• Constitutive ( house keeping) vs. Controllable genes 
• OPERON structure and its role in gene regulation 
• Regulation of Eukaryotic Gene Expression at different levels: 
• DNA methylation 
• Histon modifications(Chromatin Remodeling) 
• Increasing the number of gene copies (gene amplification) 
• Changing the rate of initiation of transcription 
• Alternate splicing 
• mRNA stability 
• Changing the rate of initiation of translation 
10/25/2014 2
Gene expression is the process by which the information 
encoded in a gene is used to direct the assembly of a protein 
molecule. 
Gene expression is explored through a study of protein structure 
and function, transcription and translation, differentiation and 
stem cells. 
It is the process by which information from a gene is used in the 
synthesis of a functional gene product. 
These products are often proteins, but in non-protein coding 
genes such as ribosomal RNA (rRNA), transfer RNA (tRNA) or 
small nuclear RNA (snRNA) genes, the product is a functional 
RNA. 
The process of gene expression is used by all known life - 
eukaryotes (including multicellular organisms), prokaryotes 
(bacteria and archaea) 
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• Regulation of gene expression includes a wide range of 
mechanisms that are used by cells to increase or decrease 
the production of specific gene products (protein or RNA). 
• Gene regulation is essential for viruses, prokaryotes and 
eukaryotes as it increases the versatility and adaptability of 
an organism by allowing the cell to express protein when 
needed. 
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• Although a functional gene product may be an RNA or a 
protein, the majority of known mechanisms regulate protein 
coding genes. 
• Any step of the gene's expression may be modulated, from 
DNA-RNA transcription to the post-translational modification 
of a protein. 
• The first discovered example of a gene regulation system was 
the lac operon, discovered by Jacques Monod, in which 
protein involved in lactose metabolism are expressed by 
E.coli only in the presence of lactose and absence of glucose. 
• Gene regulation drives the processes of cellular 
differentiation and morphogenesis, leading to the creation of 
different cell types in multicellular organisms where the 
different types of cells may possess different gene expression 
profile. 
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Genes are subunits of DNA, the information 
database of a cell that is contained inside the cell 
nucleus. 
This DNA carries the genetic blueprint that is used to 
make all the proteins the cell needs. 
Every gene contains a particular set of instructions 
that code for a specific protein 
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Constitutive ( house keeping) genes: 
 Are expressed at a fixed rate, irrespective to the cell 
condition. 
Their structure is simpler. 
Controllable genes: 
 Are expressed only as needed. Their amount may 
increase or decrease with respect to their basal level in 
different condition. 
Their structure is relatively complicated with some 
response elements 
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When the expression of genetic information is quantitatively 
increased by the presence of specific regulatory element is 
known as positive regulation. 
Element modulating positive regulation is known as activator or 
positive regulator. 
When the expression of genetic information is diminished by 
the presence of specific regulatory element is known as 
negative regulation. 
The element or molecule mediating the negative regulation is 
said to be repressor. 
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Type A response: 
Type B response: 
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1) RNA polymerase binds to DNA at promoters. 
2)Transcription initiation is regulated by proteins 
that bind to or near promoters. 
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Synthesis of the primary RNA transcript (transcription) 
Posttranscriptional modification of mRNA 
Messenger RNA degradation 
Protein synthesis ( translation ) 
Posttranslational modification of proteins 
Protein targeting & transport 
Protein degradation 10/25/2014 14
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 In prokaryotes the primary control point is the process of 
transcription initiation . 
 Different ways for regulation of gene expression in bacteria: 
 Regulation of gene expression can be done by some operon 
pathways such as 
1.lac operon. 
2.tryptophan operon. 
. 
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Transcriptional control 
Translational control 
Post translational control 
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In genetics, an operon is a functioning unit of genomic DNA 
containing a cluster of genes under the control of a single 
promoter. 
Operons occur primarily in prokaryotes but also in some 
eukaryotes. 
Operons are related to regulons, stimulons and modulons. 
An operon is made up of several structural genes arranged 
under a common promoter and regulated by a common 
operator. 
It is defined as a set of adjacent structural genes, plus the 
adjacent regulatory signals that affect transcription of the 
structural genes. 
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An operon is made up of 4 basic DNA components: 
Promoter – a nucleotide sequence that enables a gene to be 
transcribed. The promoter is recognized by RNA polymerase, which then 
initiates transcription. 
Regulator – These genes control the operator gene in cooperation 
with certain compounds called inducers and corepressors present in the 
cytoplasm. 
Operator – a segment of DNA that a repressor binds to. It is 
classically defined in the lac operon as a segment between the 
promoter and the genes of the operon. 
Structural genes – the genes that are co-regulated by the 
operon. 
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Operon regulation can be either negative or positive by induction or 
repression. 
Negative control involves the binding of a repressor to the 
operator to prevent transcription. 
In negative inducible operons, a regulatory repressor protein is normally 
bound to the operator, which prevents the transcription of the genes on 
the operon . 
If an inducer molecule is present, it binds to the repressor and changes 
its conformation so that it is unable to bind to the operator. This allows 
for expression of the operon. 
The lac operon is a negatively controlled inducible operon, where the 
inducer molecule is allolactose. 
In negative repressible operons, transcription of the operon normally 
takes place. 
The trp operon, involved in the synthesis of tryptophan (which itself acts 
as the corepressor ), is a negatively controlled repressible operon. 
10/25/2014 22
With positive control, an activator protein stimulates transcription 
by binding to DNA. 
In positive inducible operons, activator proteins are normally 
unable to bind to the pertinent DNA. 
When an inducer is bound by the activator protein, it undergoes a 
change in conformation so that it can bind to the DNA and 
activate transcription. 
In positive repressible operons, the activator proteins are 
normally bound to the pertinent DNA segment. 
However, when an inhibitor is bound by the activator, it is 
prevented from binding the DNA. 
This stops activation and transcription of the system. 
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• The lac operon of the model bacterium Escherichia coli 
was the first operon to be discovered and provides a 
typical example of operon function. 
• It consists of three adjacent structural genes, a promoter, 
a terminator, and an operator. 
• The lac operon is regulated by several factors including the 
availability of glucose and lactose. 
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Discovered in 1953 by Jacques Monod and colleagues, the trp 
operon in E. coli was the first repressible operon to be 
discovered. 
This operon contains five structural genes: 
trp E, 
trp D, 
trp C, 
trp B, and 
trp A, which encodes tryptophan synthetase. 
It also contains a promoter which binds to RNA polymerase and 
an operator which blocks transcription when bound to the 
protein synthesized by the repressor gene (trp R) that binds to 
the operator 
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Eukaryotic cells have a much larger genome 
Eukaryotes have much greater cell specialization 
Thus eukaryotic cells contain an enormous amount of DNA that 
does not program the synthesis of RNA or protein 
This requires complex organization 
In eukaryotes expression of gene into proteins can be 
controlled at various locations 
10/25/2014 35
• Synthesis of proteins is controlled right from the chromatin stage. 
• Expression of gene is controlled at many steps during the process of 
transcription and translation. 
• Description of the control points is dealt in detail in the subsequent 
slides. 
1.Transcriptional control. 
2.RNAprocessing control. 
3.RNA transport & localisation control. 
4.Translation control. 
5.mRNAdegradation control. 
6.Protein activator control. 
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Two forms of chromatin 
• Euchromatin – A lesser coiled transcriptionally active region which can 
be easily accessed by the RNA polymerases. 
• Heterochromatin – A highly condensed transcriptionally inactive region. 
The genes in this region cannot be accessed by the RNA polymerases 
for active transcription. 
• Ubiquitination: 
Ubiquitination of H2A – Transcriptional inactivation 
Ubiquitination of H2B - Transcriptional activation 
10/25/2014 38
• Histone modifications – These modifications 
make a region of gene either transcriptionally 
active or inactive. 
Acetylation 
• ↑Acetylation ----↓ Condensation of DNA ----- ↑ 
Transcription of genes in that region 
Ubiquitination 
Ubiquitination of H2A – Transcriptional inactivation 
Ubiquitination of H2B - Transcriptional activation 
10/25/2014 39
DNA methylation:is the addition or removal 
of a methyl group predominantely where 
cytosine bases occur consecutively. 
bases occur consecutively. 
DNA methylation:is the addition or removal 
of a methyl group predominantely where 
cytosine bases occur consecutively. 
bases occur consecutively. 
Methylation occurs most often in 
symmetrical CG sequences. 
10/25/2014 40
by HATs and coactivators 
leads to euchromatin 
formation 
by HDACs and 
corepressors leads to 
heterochromatin formation 
10/25/2014 41
• Eukaryotes – There are two types of promoters which are: 
• Basal promoter or core promoter -These promoters reside 
within 40bp upstream of the start site. These promoters are seen in all 
protein coding genes. 
• Upstream promoters - These promoters may lie up to 200bp 
upstream of the transcriptional initiation site. The structure of this 
promoter and the associated binding factors keeps varying from gene to 
gene. 
10/25/2014 42
Transcriptional control: 
… controlling when and how often a given gene is 
Transcribed 
Figure 6. Genes can be expressed with different efficiencies. 
Gene A is transcribed and 
translated much more efficiently than gene B. This allows the 
amount of protein A in the cell to bemuch greater than that of 
protein B. 
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Enhancers 
• Enhancers can be located upstream, downstream or 
within the gene that is transcribed 
• The binding of these enhancers with enhancer binding 
proteins (transcription factors) increases the rate of 
transcription of that gene to a greater extent. 
• Promoters are capable of initiating lower levels of 
transcription. 
• Enhancers are responsible for the cell or tissue specific 
transcription. 
• Each enhancer has its own transcription factor that it 
binds to. 
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• www.wikipedia.com 
• www.slideshare.com 
• www.webmed.com 
• www.ncbl.com 
• Some articles from internet 
• Some journals 
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REGULATION OF GENE EXPRESSION IN PROKARYOTES & EUKARYOTES

  • 2. To know and explain: • Regulation of Bacterial Gene Expression • Constitutive ( house keeping) vs. Controllable genes • OPERON structure and its role in gene regulation • Regulation of Eukaryotic Gene Expression at different levels: • DNA methylation • Histon modifications(Chromatin Remodeling) • Increasing the number of gene copies (gene amplification) • Changing the rate of initiation of transcription • Alternate splicing • mRNA stability • Changing the rate of initiation of translation 10/25/2014 2
  • 3. Gene expression is the process by which the information encoded in a gene is used to direct the assembly of a protein molecule. Gene expression is explored through a study of protein structure and function, transcription and translation, differentiation and stem cells. It is the process by which information from a gene is used in the synthesis of a functional gene product. These products are often proteins, but in non-protein coding genes such as ribosomal RNA (rRNA), transfer RNA (tRNA) or small nuclear RNA (snRNA) genes, the product is a functional RNA. The process of gene expression is used by all known life - eukaryotes (including multicellular organisms), prokaryotes (bacteria and archaea) 10/25/2014 3
  • 4. • Regulation of gene expression includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene products (protein or RNA). • Gene regulation is essential for viruses, prokaryotes and eukaryotes as it increases the versatility and adaptability of an organism by allowing the cell to express protein when needed. 10/25/2014 4
  • 5. • Although a functional gene product may be an RNA or a protein, the majority of known mechanisms regulate protein coding genes. • Any step of the gene's expression may be modulated, from DNA-RNA transcription to the post-translational modification of a protein. • The first discovered example of a gene regulation system was the lac operon, discovered by Jacques Monod, in which protein involved in lactose metabolism are expressed by E.coli only in the presence of lactose and absence of glucose. • Gene regulation drives the processes of cellular differentiation and morphogenesis, leading to the creation of different cell types in multicellular organisms where the different types of cells may possess different gene expression profile. 10/25/2014 5
  • 7. Genes are subunits of DNA, the information database of a cell that is contained inside the cell nucleus. This DNA carries the genetic blueprint that is used to make all the proteins the cell needs. Every gene contains a particular set of instructions that code for a specific protein 10/25/2014 7
  • 8. Constitutive ( house keeping) genes:  Are expressed at a fixed rate, irrespective to the cell condition. Their structure is simpler. Controllable genes:  Are expressed only as needed. Their amount may increase or decrease with respect to their basal level in different condition. Their structure is relatively complicated with some response elements 10/25/2014 8
  • 9. When the expression of genetic information is quantitatively increased by the presence of specific regulatory element is known as positive regulation. Element modulating positive regulation is known as activator or positive regulator. When the expression of genetic information is diminished by the presence of specific regulatory element is known as negative regulation. The element or molecule mediating the negative regulation is said to be repressor. 10/25/2014 9
  • 12. Type A response: Type B response: 10/25/2014 12
  • 13. 1) RNA polymerase binds to DNA at promoters. 2)Transcription initiation is regulated by proteins that bind to or near promoters. 10/25/2014 13
  • 14. Synthesis of the primary RNA transcript (transcription) Posttranscriptional modification of mRNA Messenger RNA degradation Protein synthesis ( translation ) Posttranslational modification of proteins Protein targeting & transport Protein degradation 10/25/2014 14
  • 17.  In prokaryotes the primary control point is the process of transcription initiation .  Different ways for regulation of gene expression in bacteria:  Regulation of gene expression can be done by some operon pathways such as 1.lac operon. 2.tryptophan operon. . 10/25/2014 17
  • 18. Transcriptional control Translational control Post translational control 10/25/2014 18
  • 19. In genetics, an operon is a functioning unit of genomic DNA containing a cluster of genes under the control of a single promoter. Operons occur primarily in prokaryotes but also in some eukaryotes. Operons are related to regulons, stimulons and modulons. An operon is made up of several structural genes arranged under a common promoter and regulated by a common operator. It is defined as a set of adjacent structural genes, plus the adjacent regulatory signals that affect transcription of the structural genes. 10/25/2014 19
  • 20. An operon is made up of 4 basic DNA components: Promoter – a nucleotide sequence that enables a gene to be transcribed. The promoter is recognized by RNA polymerase, which then initiates transcription. Regulator – These genes control the operator gene in cooperation with certain compounds called inducers and corepressors present in the cytoplasm. Operator – a segment of DNA that a repressor binds to. It is classically defined in the lac operon as a segment between the promoter and the genes of the operon. Structural genes – the genes that are co-regulated by the operon. 10/25/2014 20
  • 22. Operon regulation can be either negative or positive by induction or repression. Negative control involves the binding of a repressor to the operator to prevent transcription. In negative inducible operons, a regulatory repressor protein is normally bound to the operator, which prevents the transcription of the genes on the operon . If an inducer molecule is present, it binds to the repressor and changes its conformation so that it is unable to bind to the operator. This allows for expression of the operon. The lac operon is a negatively controlled inducible operon, where the inducer molecule is allolactose. In negative repressible operons, transcription of the operon normally takes place. The trp operon, involved in the synthesis of tryptophan (which itself acts as the corepressor ), is a negatively controlled repressible operon. 10/25/2014 22
  • 23. With positive control, an activator protein stimulates transcription by binding to DNA. In positive inducible operons, activator proteins are normally unable to bind to the pertinent DNA. When an inducer is bound by the activator protein, it undergoes a change in conformation so that it can bind to the DNA and activate transcription. In positive repressible operons, the activator proteins are normally bound to the pertinent DNA segment. However, when an inhibitor is bound by the activator, it is prevented from binding the DNA. This stops activation and transcription of the system. 10/25/2014 23
  • 25. • The lac operon of the model bacterium Escherichia coli was the first operon to be discovered and provides a typical example of operon function. • It consists of three adjacent structural genes, a promoter, a terminator, and an operator. • The lac operon is regulated by several factors including the availability of glucose and lactose. 10/25/2014 25
  • 30. Discovered in 1953 by Jacques Monod and colleagues, the trp operon in E. coli was the first repressible operon to be discovered. This operon contains five structural genes: trp E, trp D, trp C, trp B, and trp A, which encodes tryptophan synthetase. It also contains a promoter which binds to RNA polymerase and an operator which blocks transcription when bound to the protein synthesized by the repressor gene (trp R) that binds to the operator 10/25/2014 30
  • 35. Eukaryotic cells have a much larger genome Eukaryotes have much greater cell specialization Thus eukaryotic cells contain an enormous amount of DNA that does not program the synthesis of RNA or protein This requires complex organization In eukaryotes expression of gene into proteins can be controlled at various locations 10/25/2014 35
  • 36. • Synthesis of proteins is controlled right from the chromatin stage. • Expression of gene is controlled at many steps during the process of transcription and translation. • Description of the control points is dealt in detail in the subsequent slides. 1.Transcriptional control. 2.RNAprocessing control. 3.RNA transport & localisation control. 4.Translation control. 5.mRNAdegradation control. 6.Protein activator control. 10/25/2014 36
  • 38. Two forms of chromatin • Euchromatin – A lesser coiled transcriptionally active region which can be easily accessed by the RNA polymerases. • Heterochromatin – A highly condensed transcriptionally inactive region. The genes in this region cannot be accessed by the RNA polymerases for active transcription. • Ubiquitination: Ubiquitination of H2A – Transcriptional inactivation Ubiquitination of H2B - Transcriptional activation 10/25/2014 38
  • 39. • Histone modifications – These modifications make a region of gene either transcriptionally active or inactive. Acetylation • ↑Acetylation ----↓ Condensation of DNA ----- ↑ Transcription of genes in that region Ubiquitination Ubiquitination of H2A – Transcriptional inactivation Ubiquitination of H2B - Transcriptional activation 10/25/2014 39
  • 40. DNA methylation:is the addition or removal of a methyl group predominantely where cytosine bases occur consecutively. bases occur consecutively. DNA methylation:is the addition or removal of a methyl group predominantely where cytosine bases occur consecutively. bases occur consecutively. Methylation occurs most often in symmetrical CG sequences. 10/25/2014 40
  • 41. by HATs and coactivators leads to euchromatin formation by HDACs and corepressors leads to heterochromatin formation 10/25/2014 41
  • 42. • Eukaryotes – There are two types of promoters which are: • Basal promoter or core promoter -These promoters reside within 40bp upstream of the start site. These promoters are seen in all protein coding genes. • Upstream promoters - These promoters may lie up to 200bp upstream of the transcriptional initiation site. The structure of this promoter and the associated binding factors keeps varying from gene to gene. 10/25/2014 42
  • 43. Transcriptional control: … controlling when and how often a given gene is Transcribed Figure 6. Genes can be expressed with different efficiencies. Gene A is transcribed and translated much more efficiently than gene B. This allows the amount of protein A in the cell to bemuch greater than that of protein B. 10/25/2014 43
  • 45. Enhancers • Enhancers can be located upstream, downstream or within the gene that is transcribed • The binding of these enhancers with enhancer binding proteins (transcription factors) increases the rate of transcription of that gene to a greater extent. • Promoters are capable of initiating lower levels of transcription. • Enhancers are responsible for the cell or tissue specific transcription. • Each enhancer has its own transcription factor that it binds to. 10/25/2014 45
  • 62. • www.wikipedia.com • www.slideshare.com • www.webmed.com • www.ncbl.com • Some articles from internet • Some journals 10/25/2014 62