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Reverse Transcription

Reverse Transcription

Slide 2

Central Dogma

Central Dogma

Slide 3

Normal transcription involves synthesis of RNA from DNA.

Normal transcription involves synthesis of RNA from DNA.

Reverse transcription is the transcription of single stranded RNA into double stranded DNA

With the help of the enzyme Reverse Transcriptase.

Slide 4

Reverse Transcriptase also known as RNA directed DNA Polymerase

Reverse Transcriptase also known as RNA directed DNA Polymerase

- DNA Nucleotidyl transferase (RNA directed)

- Revertase

Reverse Transcriptase was discovered by Howard Temin and Baltimore in 1970 independently

- shared Nobel Prize in Physiology or Medicine in 1975 for their discovery.

Slide 5

Reverse transcriptase common in Retrovirus.

Reverse transcriptase common in Retrovirus.

- HIV

M-MLV (Moloney Murine Leukemia Virus)

AMV (Avian Myeloblastosis Virus)

Reverse Transcriptase enzyme includes two activity: DNA polymerase and RNase H

Slide 6

Retrovirus Replication Cycle

Retrovirus Replication Cycle

ds DNA

Provirus

host DNA

Packaging into virus ; budding

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Antiviral drug

Antiviral drug

Zidovudine

Lamivudine

Tenfovir

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Typical retrovirus has three or four genes.

Typical retrovirus has three or four genes.

Gag-pol- env

Retrovirus are called + strand because viral RNA itself code for protein products.

Reverse transcriptase enzyme code for proteins are called strand

R : Redundant or repeating seq 10-80

U5: 80 100

U3: 170-1350

Like DNA polymerase, Reverse transcriptase requires primers.

tRNA of the host is Primer

Slide 9

Mechansim Of Retrovirus Replication

Mechansim Of Retrovirus Replication

1. A Retrovirus specific cellular tRNA hybridizes with a complementary region called PBS (Primer Binding Sites)

2. Reverse Transcriptase (RT) starts at this binding site and copies RNA into a single strand of complementary DNA. A DNA segment is extended from tRNA based on the sequence of the retroviral genomic RNA

3. The viral R and U5 sequences are removed by RNase H.

LTR Long Terminal Repeat

Left LTR - Redundant sequence [R]+ 5 untranslated region (U5)

Right LTR - Redundant sequence [R]+ 3 untranslated region (U3)

Slide 10

4. First jump: DNA hybridizes with the remaining R sequence at the 3' end.

4. First jump: DNA hybridizes with the remaining R sequence at the 3' end.

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