Reverse transcriptase

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Reverse Transcriptase
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3D model of HIV reverse transcriptase

Other names: Deoxynucleoside-triphosphate:
DNA deoxynucleotidyltransferase (RNA-directed)
  • RNA-directed DNA polymerase
  • DNA nucleotidyltransferase (RNA-directed)
  • Revertase
Database Links
EC number: 2.7.7.49

In biochemistry, a reverse transcriptase, also known as RNA-dependent DNA polymerase, is a DNA polymerase enzyme that transcribes single-stranded RNA into single-stranded DNA. Normal transcription involves the synthesis of RNA from DNA; hence, reverse transcription is the reverse of this.

Reverse transcriptase was discovered by Howard Temin at the University of Wisconsin-Madison, and independently by David Baltimore in 1970. The two shared the 1975 Nobel Prize in Physiology or Medicine with Renato Dulbecco for their discovery.

Commonly used examples of reverse transcriptases include:

Function

Viruses

The enzyme is encoded and used by reverse-transcribing viruses, which use the enzyme during the process of replication. Reverse-transcribing RNA viruses, such as retroviruses, use the enzyme to reverse-transcribe their RNA genomes into DNA, which is then integrated into the host genome and replicated along with it. Reverse-transcribing DNA viruses, such as the hepadnaviruses, transcribe their genomes into an RNA intermediate and then, using reverse transcriptase, back into DNA.

Eukaryotes

Self-replicating stretches of eukaryotic genomes known as retrotransposons utilise reverse transcriptase to move from one position in the genome to another via a RNA intermediate. They are found abundantly in the genomes of plants and animals. Telomerase is another reverse transcriptase found in many eukaryotes, including humans, which carries its own RNA template; this RNA is used as a template for DNA replication[1].

Prokaryotes

Reverse transcriptases are also found in bacterial retrons, distinct sequences which code for reverse transcriptase, and are used in the synthesis of msDNA.

Structure

Reverse transcriptase enzymes include an RNA-dependent DNA polymerase and a DNA-dependent DNA polymerase, which work together to perform transcription. In addition to the transcription function, retroviral reverse transcriptases have a domain belonging to the RNase H family which is vital to their replication.

Replication fidelity

Reverse transcriptase has a high error rate when transcribing RNA into DNA since, unlike DNA polymerases, it has no proofreading ability. This high error rate allows mutations to accumulate at an accelerated rate relative to proofread forms of replication. The commercially available reverse transcriptases produced by Promega are quoted by their manuals as having error rates in the range of 1 in 17,000 bases for AMV and 1 in 30,000 bases for M-MLV[2]

Applications

The molecular structure of zidovudine (Retrovir®), a drug used to inhibit HIV reverse transcriptase

Antiviral drugs

For more details on this topic, see Reverse transcriptase inhibitor.

As HIV uses reverse transcriptase to copy its genetic material and generate new viruses (part of a retrovirus proliferation circle), specific drugs have been designed to disrupt the process and thereby suppress its growth. Collectively, these drugs are known as reverse transcriptase inhibitors and include the nucleoside and nucleotide analogues zidovudine (Retrovir®), lamivudine (Epivir®) and tenofovir (Viread®), as well as non-nucleoside inhibitors, such as nevirapine (Viramune®).

Molecular biology

For more details on this topic, see Reverse transcription polymerase chain reaction.

Reverse transcriptase is commonly used in research to apply the polymerase chain reaction technique to RNA in a technique called reverse transcription polymerase chain reaction (RT-PCR). The classical PCR technique can only be applied to DNA strands, but with the help of reverse transcriptase, RNA can be transcribed into DNA, thus making PCR analysis of RNA molecules possible. Reverse transcriptase is also used to create cDNA libraries from mRNA. The commercial availability of reverse transcriptase greatly improved knowledge in the area of molecular biology as, along with other enzymes, it allowed scientists to clone, sequence and characterise DNA.

See also

External links

References

  1. Lodish, et al, Molecular Cell Biology (2004), 5th edn, W. H. Freeman and Company, New York, ISBN 0-7167-4366-3
  2. Promega kit instruction manual (1999)
da:Revers transkriptase

de:Reverse Transkriptaseko:역전사효소 it:DNA polimerasi (RNA-dipendente) he:רוורס טרנסקריפטאז lt:Atvirkštinė transkriptazė nl:Reverse-transcriptasesv:Omvänt transkriptas ur:معکوس انتساخہ


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