bi-directional promoters

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BI-DIRECTIONAL PROMOTERS By:Sanju Sinha Date: 21-05-2014

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Page 1: BI-DIRECTIONAL PROMOTERS

BI-DIRECTIONALPROMOTERS

By:Sanju SinhaDate: 21-05-2014

Page 2: BI-DIRECTIONAL PROMOTERS

Promoters is a region of DNA that initiates the transcription of a particular gene.

What is Promoter?

Page 3: BI-DIRECTIONAL PROMOTERS

Promoter is a Important element for gene regulation.

TSS

Page 4: BI-DIRECTIONAL PROMOTERS

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.

Page 5: BI-DIRECTIONAL PROMOTERS

Promoter is a Important element for gene regulation.

TSS

Page 6: BI-DIRECTIONAL PROMOTERS

On Basis of Directions they can transcribe, Promoters can be classified into two sub-classes-

1.Unidirectional 2.Bi-Directional*

Page 7: BI-DIRECTIONAL PROMOTERS

Definition-Bidirectional promoters are short (<1 kbp), intergenic regions of DNA between the 5‘ ends of the genes in a bidirectional gene pair.

Page 8: BI-DIRECTIONAL PROMOTERS

Head-To-Head Fashion Alignment

Page 9: BI-DIRECTIONAL PROMOTERS

1000 BP1200 BP1500 BP

So Lets Increase This Window

10000 BP12000 BP15000 BPVS

Page 10: BI-DIRECTIONAL PROMOTERS

0 200 400 600 800 1000 1200 1400 16000

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f(x) = 1251.38499631088 ln(x) − 3205.50840193062

Series1 Logarithmic (Series1)

Page 11: BI-DIRECTIONAL PROMOTERS

1000 BP1200 BP1500 BP

So Lets Increase This Window

10000 BP12000 BP15000 BPVS

Page 12: BI-DIRECTIONAL PROMOTERS

0 5000 10000 15000 20000 250000

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Chart Title

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what is the promoter length Distribution then?

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Promoter Length Histogram (window =1500)

100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 More0

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0 5000 10000 15000 20000 250000

2000400060008000

1000012000140001600018000

Chart Title

0 200 400 600 800 1000 1200 1400 16000

1000

2000

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f(x) = 1251.38499631088 ln(x) − 3205.50840193062

Series1Logarithmic (Series1)

100300

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0100200300400500600700800

Frequency

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On Basis of these Results:

Mean Promoter length is : 335 Median of Promoter length is: 189

Page 16: BI-DIRECTIONAL PROMOTERS

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.

Page 17: BI-DIRECTIONAL PROMOTERS

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.

Page 18: BI-DIRECTIONAL PROMOTERS

1. All Further Data is beingtaken from a review paper.

2. All the sources and platforms are mentioned on last slide.

PART - 2

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Is it possible to identify consistent pattern that distinguish Bidirectional and Unidirectional ???

What to Look for …..

Page 20: BI-DIRECTIONAL PROMOTERS

GC content

INR

TATA Box

BRE

DPE

CCAAT

Page 21: BI-DIRECTIONAL PROMOTERS

Location of different elements of PROMOTER(lacking TATA)

Page 22: BI-DIRECTIONAL PROMOTERS

GC content

INR

TATA Box

BRE

DPE

CCAAT

Page 23: BI-DIRECTIONAL PROMOTERS

Statistical Results of GC content:Average GC content percentage

Bidirectional: 66%

Unidirectional: 53%

Page 24: BI-DIRECTIONAL PROMOTERS

• Bidirectional: 66%• Unidirectional: 53%GC content

INR

TATA Box

BRE

DPE

CCAAT

Page 25: BI-DIRECTIONAL PROMOTERS

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%

Page 26: BI-DIRECTIONAL PROMOTERS

• Bidirectional: 66%• Unidirectional: 53%GC content• Bidirectional: 25.3%• Unidirectional: 30.8%INR

TATA Box

BRE

DPE

CCAAT

Page 27: BI-DIRECTIONAL PROMOTERS

TATA box:

Bidirectional: Most of them Lacks.

Unidirectional: Comparatively high.

• Located at -30 both in Unidirec and Bi-direc Promoters

Page 28: BI-DIRECTIONAL PROMOTERS

• Bidirectional: 66%• Unidirectional: 53%GC content• Bidirectional: 25.3%• Unidirectional: 30.8%INR• Bidirectional: Lacks Mostly(No

data)• Unidirectional: Have more

frequently

TATA Box

BRE

DPE

CCAAT

Page 29: BI-DIRECTIONAL PROMOTERS

BRE(B-recognition element)• Located directly in front of TATA• TFIIB recognizes it and binds.

Bidirectional: 16.5%

Unidirectional: 11%

Page 30: BI-DIRECTIONAL PROMOTERS

• Bidirectional: 66%• Unidirectional: 53%GC content• Bidirectional: 25.3%• Unidirectional: 30.8%INR• Bidirectional: Lacks mostly(No

Data)• Unidirectional: Have more

frequently

TATA Box • Bidirectional: 16.5%• Unidirectional: 11.1%BRE

DPE

CCAAT

Page 31: BI-DIRECTIONAL PROMOTERS

DPE(Downstream Promoter Element)• Located at +30 position• Binds to common transcription

factor(TFIID) in absence of TATA

Bidirectional: 46.6%

Unidirectional: 50.6%

Page 32: BI-DIRECTIONAL PROMOTERS

• Bidirectional: 66%• Unidirectional: 53%GC content• Bidirectional: 25.3%• Unidirectional: 30.8%INR• Bidirectional: Lacks mostly(No

Data)• Unidirectional: Have more

frequently

TATA Box • Bidirectional: 16.5%• Unidirectional: 11%BRE• Bidirectional: 46.6%• Unidirectional: 50.6%DPE

CCAAT

Page 33: BI-DIRECTIONAL PROMOTERS

CCAAT box:• Located at 75-80 BP before TSS• signals the binding site for the

RNA transcription factor

Bidirectional: 12.9%

Unidirectional: 6.9%

Page 34: BI-DIRECTIONAL PROMOTERS

• Bidirectional: 66%• Unidirectional: 53%GC content• Bidirectional: 25.3%• Unidirectional: 30.8%INR• Bidirectional: Lacks mostly(No

Data)• Unidirectional: Have more

frequently

TATA Box • Bidirectional: 16.5%• Unidirectional: 11.1%BRE• Bidirectional: 46.6%• Unidirectional: 50.6%DPE• Bidirectional: 12.9%• Unidirectional: 6.9%CCAAT

Page 35: BI-DIRECTIONAL PROMOTERS

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.

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Bi-Directional promoters enrich with following specific Binding sites of TF.

• GABPA• MYC• E2F1• E2F4• Nrf1• YY1• NFY• SP1

Page 37: BI-DIRECTIONAL PROMOTERS

PART- 3Follow the tunnel

So the first thing is- GC CONTENT

Page 38: BI-DIRECTIONAL PROMOTERS

Lets check the GC content-

Wait..Wait..Wait..

Where’s the length of Unidirectional Promoters??

Page 39: BI-DIRECTIONAL 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

Page 40: BI-DIRECTIONAL PROMOTERS

Comparison of GC content between

Unidirectional (Mean Length)Bidirectional VS Unidirectional (Median Length)

Unidirectional (1000bp Length)

Page 41: BI-DIRECTIONAL PROMOTERS

GC content Distribution- Unidirectional_MEAN

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More0

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AVERAGE: 54.6%

Page 42: BI-DIRECTIONAL PROMOTERS

GC content Distribution- Unidirectional_MEDIAN

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More0

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AVERAGE: 56.9%

Page 43: BI-DIRECTIONAL PROMOTERS

GC content Distribution- Unidirectional_1000 BP

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More0

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AVERAGE: 49.7%

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Comparison of GC content between

Unidirectional (Mean Length)Bidirectional VS Unidirectional (Median Length)

Unidirectional (1000bp Length)

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GC content Distribution- BIDIRECTIONAL

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 More0

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AVERAGE: 64%

Page 46: BI-DIRECTIONAL PROMOTERS

Statistical Conclusion:

Unidirectional: 53.7%

Bidirectional: 64%

Average GC content :

Page 47: BI-DIRECTIONAL PROMOTERS

NOTE**DATA SOURCE and Platforms1. 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: [email protected]

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.

Page 48: BI-DIRECTIONAL PROMOTERS

THANKS