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BI-DIRECTIONAL
PROMOTERS
By:Sanju Sinha
Date: 21-05-2014
Promoters is a region of DNA
that initiates the transcription
of a particular gene.
What is Promoter?
Promoter is a Important element for gene regulation.
TSS
Assumptions
• Promoters are
Usually
conceptualized as
upstream of the
sequences they
promote.
Facts
• Scientist do not really
know in which
direction promoters
usually transcribe or
if they only
transcribe in one
direction or not.
Present-Research
• Their Directions
possibilities and
parts of
promoter which
plays role in
deciding direction.
Promoter is a Important element for gene regulation.
TSS
On Basis of Directions they can
transcribe, Promoters can be
classified into two sub-classes-
1.Unidirectional
2.Bi-Directional*
Definition-
Bidirectional promoters
are short (<1 kbp), intergenic
regions of DNA between the
5‘ ends of the genes in a
bidirectional gene pair.
Head-To-Head Fashion Alignment
1000 BP
1200 BP
1500 BP
So Lets Increase This Window
10000 BP
12000 BP
15000 BPVS
0 200 400 600 800 1000 1200 1400 1600
0
1000
2000
3000
4000
5000
6000
7000
Series1 Log. (Series1)
1000 BP
1200 BP
1500 BP
So Lets Increase This Window
10000 BP
12000 BP
15000 BPVS
0
2000
4000
6000
8000
10000
12000
14000
16000
18000
0 5000 10000 15000 20000 25000
Chart Title
what is the promoter length Distribution then?
Promoter Length Histogram (window =1500)
0
100
200
300
400
500
600
700
800
100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 More
Frequency
Bin
Frequency
0
5000
10000
15000
20000
0 5000 10000 15000 20000 25000
Chart Title
0 500 1000 1500 2000
0
2000
4000
6000
8000
Series1 Log. (Series1)
0
100
200
300
400
500
600
700
800
100
300
500
700
900
1100
1300
1500
Frequency
Bin
Frequency
On Basis of these Results:
Mean Promoter length is : 335 Median of Promoter length is: 189
Conclusion we can draw:
• Clusters near the gene starting position in
range of 189.
• The probability of occurrence of another
gene at a distance from one gene first
increases exponentially till 335 and then
decreases and then saturates tending to
constant*.
*Not sure as second differentiation is still positive and can even change its concavity.
Visions and Logics to verify data:
• Making a artificial gene distribution
like system.
• A Cyber-Refgene.txt file.
• Using the same tunnel and get the
distribution.
• Comparing the Distribution.
1. All Further Data is being
taken from a review paper.
2. All the sources and platforms
are mentioned on last slide.
PART - 2
Is it possible to identify consistent
pattern that distinguish
Bidirectional and Unidirectional ???
What to Look for …..
GC content
INR
TATA Box
BRE
DPE
CCAAT
Location of different elements of PROMOTER(lacking TATA)
GC content
INR
TATA Box
BRE
DPE
CCAAT
Statistical Results of GC content:
Average GC content percentage
Bidirectional: 66%
Unidirectional: 53%
• Bidirectional: 66%
• Unidirectional: 53%GC content
INR
TATA Box
BRE
DPE
CCAAT
INR-Initiator element
• Functionally similar to TATA box.
• Accurate transcription initiation ,
INR btw -3 to +5 is necessary.
• Increases the strength of TATA
containing promoters.
Bidirectional: 25.3%
Unidirectional: 30.8%
• Bidirectional: 66%
• Unidirectional: 53%GC content
• Bidirectional: 25.3%
• Unidirectional: 30.8%INR
TATA Box
BRE
DPE
CCAAT
TATA box:
Bidirectional: Most of them Lacks.
Unidirectional: Comparatively high.
• Located at -30 both in Unidirec and
Bi-direc Promoters
• Bidirectional: 66%
• Unidirectional: 53%GC content
• Bidirectional: 25.3%
• Unidirectional: 30.8%INR
• Bidirectional: Lacks Mostly(No data)
• Unidirectional: Have more frequentlyTATA Box
BRE
DPE
CCAAT
BRE(B-recognition element)
• Located directly in front of TATA
• TFIIB recognizes it and binds.
Bidirectional: 16.5%
Unidirectional: 11%
• Bidirectional: 66%
• Unidirectional: 53%GC content
• Bidirectional: 25.3%
• Unidirectional: 30.8%INR
• Bidirectional: Lacks mostly(No Data)
• Unidirectional: Have more frequentlyTATA Box
• Bidirectional: 16.5%
• Unidirectional: 11.1%BRE
DPE
CCAAT
DPE(Downstream Promoter Element)
• Located at +30 position
• Binds to common transcription
factor(TFIID) in absence of TATA
Bidirectional: 46.6%
Unidirectional: 50.6%
• Bidirectional: 66%
• Unidirectional: 53%GC content
• Bidirectional: 25.3%
• Unidirectional: 30.8%INR
• Bidirectional: Lacks mostly(No Data)
• Unidirectional: Have more frequentlyTATA Box
• Bidirectional: 16.5%
• Unidirectional: 11%BRE
• Bidirectional: 46.6%
• Unidirectional: 50.6%DPE
CCAAT
CCAAT box:
• Located at 75-80 BP before TSS
• signals the binding site for the
RNA transcription factor
Bidirectional: 12.9%
Unidirectional: 6.9%
• Bidirectional: 66%
• Unidirectional: 53%GC content
• Bidirectional: 25.3%
• Unidirectional: 30.8%INR
• Bidirectional: Lacks mostly(No Data)
• Unidirectional: Have more frequentlyTATA Box
• Bidirectional: 16.5%
• Unidirectional: 11.1%BRE
• Bidirectional: 46.6%
• Unidirectional: 50.6%DPE
• Bidirectional: 12.9%
• Unidirectional: 6.9%CCAAT
CpG islands:
The CpG sites or CG sites are regions of DNA where
a cytosine nucleotide occurs next to a guanine
Source:Trinklein, N. D., Aldred, S. H., Hartman, S. J., Schroeder, D. I., Otillar, R. P., and Myers, R. M. (2004) Genome Res.,14, 6266.
• 77% B-DP located in CpG islands
compared with 38% of U-DP.
• 90% B-DP located in CpG islands
compared with 45% of U-DP.
Source: Yang, M. Q., and Elnitski, L. L. (2008) BMC Genom., 9 (Suppl. 2), S3.
Bi-Directional promoters enrich with
following specific Binding sites of TF.
• GABPA
• MYC
• E2F1
• E2F4
• Nrf1
• YY1
• NFY
• SP1
PART- 3
Follow the tunnel
So the first thing is
- GC CONTENT
Lets check the GC content-
Wait..Wait..Wait..
Where’s the length of
Unidirectional Promoters??
No, We Don’t . But we
have some Important
values which can help us.
1. Mean length of Bidirectional Promoter.
2. Median Length of Bidirectional Promoters.
3. We Know in Paper they take 1000BP
Comparison of GC content between
Unidirectional (Mean Length)
Bidirectional VS Unidirectional (Median Length)
Unidirectional (1000bp Length)
GC content Distribution- Unidirectional_MEAN
0
500
1000
1500
2000
2500
3000
3500
4000
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More
Frequency
Bin
Histogram
Frequency
AVERAGE: 54.6%
GC content Distribution- Unidirectional_MEDIAN
0
500
1000
1500
2000
2500
3000
3500
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More
Frequency
Bin
Histogram
Frequency
AVERAGE: 56.9%
GC content Distribution- Unidirectional_1000 BP
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More
Frequency
Bin
Histogram
Frequency
AVERAGE: 49.7%
Comparison of GC content between
Unidirectional (Mean Length)
Bidirectional VS Unidirectional (Median Length)
Unidirectional (1000bp Length)
GC content Distribution- BIDIRECTIONAL
0
100
200
300
400
500
600
700
800
900
1000
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More
Frequency
Bin
Histogram
Frequency
AVERAGE: 64%
Statistical Conclusion:
Unidirectional: 53.7%
Bidirectional: 64%
Average GC content :
NOTE**DATA SOURCE and Platforms
1. All the Data mentioned in Slides 17-36 are taken from Review:
Bidirectional Promoters in the Transcription of Mammalian Genomes.
A. S. Orekhova and P. M. Rubtsov*
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, 119991
Moscow, Russia; fax: (499) 1351405; Email: rubtsov@eimb.ru
2. All other data in these presentation belong to Sanju Sinha and he have all rights on those.
Any copying without mentioning the relevance source shall be considered as plagiarism.
3.twoBitToFa on linux platform is being used to done the calculations.
4. All coding is being done via Python language.
THANKS

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BI-DIRECTIONAL PROMOTERS

  • 2. Promoters is a region of DNA that initiates the transcription of a particular gene. What is Promoter?
  • 3. Promoter is a Important element for gene regulation. TSS
  • 4. Assumptions • Promoters are Usually conceptualized as upstream of the sequences they promote. Facts • Scientist do not really know in which direction promoters usually transcribe or if they only transcribe in one direction or not. Present-Research • Their Directions possibilities and parts of promoter which plays role in deciding direction.
  • 5. Promoter is a Important element for gene regulation. TSS
  • 6. On Basis of Directions they can transcribe, Promoters can be classified into two sub-classes- 1.Unidirectional 2.Bi-Directional*
  • 7. Definition- Bidirectional promoters are short (<1 kbp), intergenic regions of DNA between the 5‘ ends of the genes in a bidirectional gene pair.
  • 9. 1000 BP 1200 BP 1500 BP So Lets Increase This Window 10000 BP 12000 BP 15000 BPVS
  • 10. 0 200 400 600 800 1000 1200 1400 1600 0 1000 2000 3000 4000 5000 6000 7000 Series1 Log. (Series1)
  • 11. 1000 BP 1200 BP 1500 BP So Lets Increase This Window 10000 BP 12000 BP 15000 BPVS
  • 13. what is the promoter length Distribution then?
  • 14. Promoter Length Histogram (window =1500) 0 100 200 300 400 500 600 700 800 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 More Frequency Bin Frequency
  • 15. 0 5000 10000 15000 20000 0 5000 10000 15000 20000 25000 Chart Title 0 500 1000 1500 2000 0 2000 4000 6000 8000 Series1 Log. (Series1) 0 100 200 300 400 500 600 700 800 100 300 500 700 900 1100 1300 1500 Frequency Bin Frequency On Basis of these Results: Mean Promoter length is : 335 Median of Promoter length is: 189
  • 16. Conclusion we can draw: • Clusters near the gene starting position in range of 189. • The probability of occurrence of another gene at a distance from one gene first increases exponentially till 335 and then decreases and then saturates tending to constant*. *Not sure as second differentiation is still positive and can even change its concavity.
  • 17. Visions and Logics to verify data: • Making a artificial gene distribution like system. • A Cyber-Refgene.txt file. • Using the same tunnel and get the distribution. • Comparing the Distribution.
  • 18. 1. All Further Data is being taken from a review paper. 2. All the sources and platforms are mentioned on last slide. PART - 2
  • 19. Is it possible to identify consistent pattern that distinguish Bidirectional and Unidirectional ??? What to Look for …..
  • 21. Location of different elements of PROMOTER(lacking TATA)
  • 23. Statistical Results of GC content: Average GC content percentage Bidirectional: 66% Unidirectional: 53%
  • 24. • Bidirectional: 66% • Unidirectional: 53%GC content INR TATA Box BRE DPE CCAAT
  • 25. INR-Initiator element • Functionally similar to TATA box. • Accurate transcription initiation , INR btw -3 to +5 is necessary. • Increases the strength of TATA containing promoters. Bidirectional: 25.3% Unidirectional: 30.8%
  • 26. • Bidirectional: 66% • Unidirectional: 53%GC content • Bidirectional: 25.3% • Unidirectional: 30.8%INR TATA Box BRE DPE CCAAT
  • 27. TATA box: Bidirectional: Most of them Lacks. Unidirectional: Comparatively high. • Located at -30 both in Unidirec and Bi-direc Promoters
  • 28. • Bidirectional: 66% • Unidirectional: 53%GC content • Bidirectional: 25.3% • Unidirectional: 30.8%INR • Bidirectional: Lacks Mostly(No data) • Unidirectional: Have more frequentlyTATA Box BRE DPE CCAAT
  • 29. BRE(B-recognition element) • Located directly in front of TATA • TFIIB recognizes it and binds. Bidirectional: 16.5% Unidirectional: 11%
  • 30. • Bidirectional: 66% • Unidirectional: 53%GC content • Bidirectional: 25.3% • Unidirectional: 30.8%INR • Bidirectional: Lacks mostly(No Data) • Unidirectional: Have more frequentlyTATA Box • Bidirectional: 16.5% • Unidirectional: 11.1%BRE DPE CCAAT
  • 31. DPE(Downstream Promoter Element) • Located at +30 position • Binds to common transcription factor(TFIID) in absence of TATA Bidirectional: 46.6% Unidirectional: 50.6%
  • 32. • Bidirectional: 66% • Unidirectional: 53%GC content • Bidirectional: 25.3% • Unidirectional: 30.8%INR • Bidirectional: Lacks mostly(No Data) • Unidirectional: Have more frequentlyTATA Box • Bidirectional: 16.5% • Unidirectional: 11%BRE • Bidirectional: 46.6% • Unidirectional: 50.6%DPE CCAAT
  • 33. CCAAT box: • Located at 75-80 BP before TSS • signals the binding site for the RNA transcription factor Bidirectional: 12.9% Unidirectional: 6.9%
  • 34. • Bidirectional: 66% • Unidirectional: 53%GC content • Bidirectional: 25.3% • Unidirectional: 30.8%INR • Bidirectional: Lacks mostly(No Data) • Unidirectional: Have more frequentlyTATA Box • Bidirectional: 16.5% • Unidirectional: 11.1%BRE • Bidirectional: 46.6% • Unidirectional: 50.6%DPE • Bidirectional: 12.9% • Unidirectional: 6.9%CCAAT
  • 35. CpG islands: The CpG sites or CG sites are regions of DNA where a cytosine nucleotide occurs next to a guanine Source:Trinklein, N. D., Aldred, S. H., Hartman, S. J., Schroeder, D. I., Otillar, R. P., and Myers, R. M. (2004) Genome Res.,14, 6266. • 77% B-DP located in CpG islands compared with 38% of U-DP. • 90% B-DP located in CpG islands compared with 45% of U-DP. Source: Yang, M. Q., and Elnitski, L. L. (2008) BMC Genom., 9 (Suppl. 2), S3.
  • 36. Bi-Directional promoters enrich with following specific Binding sites of TF. • GABPA • MYC • E2F1 • E2F4 • Nrf1 • YY1 • NFY • SP1
  • 37. PART- 3 Follow the tunnel So the first thing is - GC CONTENT
  • 38. Lets check the GC content- Wait..Wait..Wait.. Where’s the length of Unidirectional Promoters??
  • 39. No, We Don’t . But we have some Important values which can help us. 1. Mean length of Bidirectional Promoter. 2. Median Length of Bidirectional Promoters. 3. We Know in Paper they take 1000BP
  • 40. Comparison of GC content between Unidirectional (Mean Length) Bidirectional VS Unidirectional (Median Length) Unidirectional (1000bp Length)
  • 41. GC content Distribution- Unidirectional_MEAN 0 500 1000 1500 2000 2500 3000 3500 4000 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More Frequency Bin Histogram Frequency AVERAGE: 54.6%
  • 42. GC content Distribution- Unidirectional_MEDIAN 0 500 1000 1500 2000 2500 3000 3500 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More Frequency Bin Histogram Frequency AVERAGE: 56.9%
  • 43. GC content Distribution- Unidirectional_1000 BP 0 500 1000 1500 2000 2500 3000 3500 4000 4500 5000 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More Frequency Bin Histogram Frequency AVERAGE: 49.7%
  • 44. Comparison of GC content between Unidirectional (Mean Length) Bidirectional VS Unidirectional (Median Length) Unidirectional (1000bp Length)
  • 45. GC content Distribution- BIDIRECTIONAL 0 100 200 300 400 500 600 700 800 900 1000 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More Frequency Bin Histogram Frequency AVERAGE: 64%
  • 47. NOTE**DATA SOURCE and Platforms 1. All the Data mentioned in Slides 17-36 are taken from Review: Bidirectional Promoters in the Transcription of Mammalian Genomes. A. S. Orekhova and P. M. Rubtsov* Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, 119991 Moscow, Russia; fax: (499) 1351405; Email: rubtsov@eimb.ru 2. All other data in these presentation belong to Sanju Sinha and he have all rights on those. Any copying without mentioning the relevance source shall be considered as plagiarism. 3.twoBitToFa on linux platform is being used to done the calculations. 4. All coding is being done via Python language.