Reverse Transcription !

Kyle Foster

Scientific Writing section 2

Dr. Bruce

26 February 2018

Reverse Transcription
Introduction

Reverse Transcription is a biological process in which ribonucleic acid (RNA) is copied

into chromosomal deoxyribonucleic acid (cDNA). It was long thought that such a process did not

occur in biological systems, because it contradicted the

central dogma of molecular biology. The central dogma of

molecular biology is a scientific theory that an

organism’s DNA is converted to mRNA by RNA

polymerase, which is then translated to a protein by

ribosomes. Proteins carry out most of the functions in

biological systems. The understanding of this

information flow is important to know before going

in depth of how reverse transcription works, because Figure 1. Central Dogma of Molecular
Biochemistry with Enzymes (Horspool, 2008)
reverse transcription takes advantage of the

molecular machinery used in this central dogma.

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A Brief Introduction to the Nucleic Acids

DNA and RNA are both nucleic acids, but differ in subtle yet important ways. DNA

stands for deoxyribonucleic acid and RNA stands for ribonucleic acids. One of the differences

between the two nucleic acids is in the ribose sugar backbone. Figure 2 depicts a ribose sugar

which is composed of a 5 membered ring with 4 carbon atoms represented at the corners of the

hexagon and oxygen. Focusing only on the number 2 carbon, you can see that in DNA, the

Figure 2. Ribose deoxyribose (username: Boumphreyfr, 2009)

carbon is attached to a hydrogen and in RNA, that carbon is bonded to an OH. There is also a

subtle difference between the bases that DNA and RNA

attach to on the carbon sugar backbone on carbon 1.

Figure 3 illustrates a segment of DNA where the

yellow strand is the ribose sugar backbone with bases

inbetwwe Both DNA and RNA can have Cytosine (C), Guanine (G) and

Adenine (A) bases, but only DNA can have thymine

(T) while only RNA can have uracil (U). When Figure 3. Diagram
showing a double
looking at a double strand of DNA, complementary helix of a
chromosome
CRUK 065 (Cancer
Research UK,
Reverse Transcription !3

matching is observed between the two strands. DNA is double stranded due to the hydrogen

bonding between complementary bases. This complementary matching allows for DNA

replication and transcription. The nucleic acids always line up as follows: A always matches with

T, and G always matches with C. A strand of nucleotides that codes for a protein is called a gene.

Organisms have thousands of genes, the aggregate of which is called the genome.

The Process of Transcription

Transcription occurs one gene at a time by a protein enzyme called RNA polymerase. RNA

polymerase is a very big and complex protein that is able to unwind DNA and synthesize a

complimentary strand of messenger RNA using the complementary rules stated earlier, but when

the RNA polymerase reads an A, instead of

adding a T, it adds a U. Recall that RNA does

not have T as a base. All other base paring

remain the same. Figure 4 illustrates a

simplified example of the process of

transcription. Another important

Figure 4. Process of transcription (NHS National
Genetics and Genomics Education Centre, 2014) distinction between DNA and RNA is that

RNA is usually single stranded while DNA

is double stranded. This RNA strand can now go throughout the cell and be translated by

ribosomes to form functional proteins.

The Process of Reverse Transcription

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Reverse transcription is the process of taking RNA and converting it into chromosomal

DNA. The process of reverse transcription requires two proteins that usually come with a viral

(foreign) DNA or RNA. Reverse transcriptase is able to create DNA strands using either an RNA

or a DNA template. When the reverse transcriptase synthesizes the new DNA, a DNA/RNA

double helix is formed. This structure is unstable, so it breaks apart into 2 separate strands.

Reverse transcriptase is able to use the newly synthesized DNA strand as a template to make

another strand of DNA, which then makes a stable double stranded DNA molecule that can

implant itself into the host cell’s chromosomal DNA. The newly synthesized double stranded

DNA is integrated into the host genome by integrase, a protein that is usually encoded by the

viral DNA. (Alberts et al, 2014).

Reverse Transcription in Retroviruses

Retroviruses use reverse transcription

to reproduce. Viruses are simple

biological structures which are

composed of RNA or DNA

surrounded by a protein coat. The

protein coat can also hold proteins

from previous host. Some of these

proteins help with viral infectivity and

others have no signifiant effect

(Kishimoto, 2017). Regardless,

viruses have very little molecular

Figure 5. HIV Virus Replication Cycle (NIAID, 2010)

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machinery and reproduce using host cell machinery through processes of DNA replication,

translation and reverse transcription. The benefit of this process is that the viral DNA will now

replicate anytime the cell replicates its own DNA. Viral DNA can remain dormant for years, but

once the viral DNA is transcribed, the cell’s many mRNAs are translated into proteins that make

new viruses which can then go on to infect other cells.

Being infected with HIV today is much better than 20 years ago because we have

developed drugs that stop the process of reverse transcription. Many people are alive today due

to the understanding of this research. Reverse transcription is also useful in the laboratory for

many different protocols such as RNA sequencing.

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Works Cited

Alberts, B., Bray, D., Hopkin, K., Johnson, A., Lewis, J., Raff, M., . . . Walter, P. (2014).

Essential cell biology. New York, NY: Garland Science.

Boumphreyfr. (2009, July 1). Ribose deoxyribose [Digital image]. Retrieved February 25, 2018,

from https://commons.wikimedia.org/wiki/File:Ribose_deoxyribose.png

Cancer Research UK. (2014, July 30). Diagram showing a double helix of a chromosome CRUK

[Digital image]. Retrieved February 25, 2018, from https://commons.wikimedia.org/wiki/

File:Diagram_showing_a_double_helix_of_a_chromosome_CRUK_065.svg

Horspool, D. (2008, November 28). Central Dogma of Molecular Biochemistry with Enzymes

[Digital image]. Retrieved February 25, 2018, from https://commons.wikimedia.org/wiki/

File:Central_Dogma_of_Molecular_Biochemistry_with_Enzymes.jpg

Kishimoto, N., Iga, N., Yamamoto, K., Takamune, N., & Misumi, S. (2017). Virion-incorporated

alpha-enolase suppresses the early stage of HIV-1 reverse transcription. Biochemical and

Biophysical Research Communications, 484(2), 278-284. doi:10.1016/j.bbrc.2017.01.096

NHS National Genetics and Genomics Education Centre. (2014, March 11). Process of

transcription [Digital image]. Retrieved February 25, 2018, from https://

commons.wikimedia.org/wiki/File:Process_of_transcription_(13080846733).jpg

NIAID. (2010, October 6). HIV Virus Replication Cycle [Digital image]. Retrieved February 25,

2018, from https://www.flickr.com/photos/niaid/5057022555

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Post-write

I used several methods to convey the complex idea of reverse transcription. Much of

molecular biology is the interaction of molecules so I used lots of images to illustrate the

characters that interact with each other. I also made made distinctions between similar molecules

such as DNA and RNA and carefully explain what makes them different. I used an example of a

retrovirus by talking about how HIV infects a cell and uses reverse transcription to replicate

which also incorporated reverse transcription into a real relevant scenario.