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Post-Transcriptional Regulation

The inside of a cell can be a battlefield between foreign material and the cell itself. Viruses can release their RNA into human cells during an infection and human cells have mechanisms in place to protect against these viruses. 

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Post-Transcriptional Regulation

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The inside of a cell can be a battlefield between foreign material and the cell itself. Viruses can release their RNA into human cells during an infection and human cells have mechanisms in place to protect against these viruses.

However, the protection mechanism is not very specific. Cells will target any single-stranded RNA for degradation which poses a big problem when humans also synthesize RNA to make proteins.

To circumvent this problem, human cells have included post-transcriptional modifications to human-synthesized RNA to protect against the harsh environment of the cell. So, if you are interested in learning more about post-transcriptional regulation, keep reading!

  • First, we will talk about the post-transcriptional control of gene expression
  • Then, we will look at the types of post-transcriptional regulation.
  • After, we will explore post-transcriptional regulation in eukaryotes.
  • Lastly, we will look at the post-transcriptional regulation in prokaryotes.

Post-Transcriptional Control of Gene Expression

Recall the Central Dogma by which instructions in the form of DNA in the cell are made into RNA before eventually being made into proteins (Fig. 1).

During transcription, DNA is turned into heterogeneous nuclear RNA (hnRNA). Following transcription, a series of enzyme-catalyzed modifications called post-transcriptional modifications occur to convert hnRNA into functional messenger RNA (mRNA).

hnRNA become mRNA following post-transcriptional modifications

In eukaryotic cells, both transcription and post-transcriptional modifications occur in the nucleus while translation occurs in the cytoplasm. As viruses and bacteria use single-stranded RNA to replicate inside the cytoplasm, eukaryotic cells have developed mechanisms to degrade single-stranded RNA as a protective mechanism.

If hnRNAs without post-transcriptional modifications are released into the cytoplasm, they would be degraded by the cell's own degradation machinery. Post-transcriptional modifications in the nucleus allow mRNA to avoid self-degradation and survive in the harsh environment of the cytoplasm.

In contrast, prokaryotic cells do not have organelles or a nucleus. Therefore, both transcription and translation occur in the cytoplasm. To prevent RNA from being degraded in the cytoplasm, prokaryotes begin translation before transcription has been completed.

Therefore, prokaryotic cells do not have post-transcriptional modifications, instead, prokaryotes have alternative mechanisms to validate mRNA quality and prevent viral replication.

Types of Post-Transcriptional Regulation

There are three post-transcriptional modifications that must occur to the hnRNA before it can be released into the cytoplasm as a mature mRNA:

  1. Addition of a 5' GTP cap.
  2. Addition of a 3' poly-A tail.
  3. Removal of introns and retention of exons.

The details of each modification will be discussed in the next section.

Post-Transcriptional Regulation in Eukaryotes

Addition of a 5' GTP cap

Recall that RNA and DNA have directionality with a 5' end and a 3' end. At the end of the 5' end of hnRNA, a 7-methylguanylate-triphosphate cap also known as a 5' GTP cap is added. The 5' GTP cap is a modified guanine nucleotide with an additional methyl group at the 7th position of guanine.

The addition of the 5' GTP cap is catalyzed by three enzymes which are part of the CAP-binding complex. The CAP-binding complex will connect the 5' carbon of the GTP cap to the 5' carbon of the first nucleotide to create a 5'-to-5' triphosphate bridge.

There are three overall functions of the 5' GTP cap:

  1. To aid in RNA processing and export of the RNA from the nucleus.

  2. To act as a marker to orient the mRNA during translation in the cytoplasm.

  3. To protect the mRNA from degradation.

Addition of a 3' poly-A tail

The polyadenosyl (poly-A) tail is added to the 3' end of hnRNA to protect the RNA transcript from rapid degradation in the cytoplasm.

Once any single-stranded RNA strand enters the cytoplasm, it will be rapidly degraded by the cell's degradation machinery known as 3'-exonucleases. The 3' exonucleases rapidly degrade single-stranded RNA beginning from its 3' end as a protective mechanism against viral and bacterial RNA transcripts. Once the mRNA is released into the cytoplasm, it too will be degraded by 3'-exonucleases.

However, the poly-A tail acts as a buffer that will be degraded before the coding information on the mRNA. The longer the poly-A tail, the longer that the mRNA transcript can survive in the cytoplasm. The poly-A tail is also critical for the export of mature mRNA from the nucleus into the cytoplasm.

The addition of the 5' GTP cap and the 3' poly-A tail is illustrated in Figure 2 below.

Removal of Introns

Recall that eukaryotic genes are made of coding sequences known as exons and non-coding sequences known as introns. Often introns are spaced between exons, therefore, following the transcription of DNA into RNA, the introns must be removed and the exons must be reassembled into one mRNA strand (Fig. 3).

The process to remove introns is known as splicing. Splicing is performed by a complex called the spliceosome which itself is made up of non-coding RNA called small nuclear RNA (snRNA) and proteins called small nuclear ribonucleoprotein (snRNPs).

Together, they make up the spliceosome which will recognize the boundaries of the intron and excise them from the RNA to be degraded. The final product is an RNA transcript that is made up of only exons.

Some eukaryotic genes, however, are made up of unique combinations of exons. This means that not all exons are transcribed into a mature mRNA.

In Figure 4, different exons are included in the final RNA transcript. On the left, exon #2 but not exon #3 may be included in mRNA #1 while the opposite may be true for mRNA #2. This selective inclusion of exons is known as alternative splicing. Alternative splicing allows for different forms of the protein to be made from the same DNA sequence.

Post-Transcriptional Regulation Examples

Recall that all cells in the human body have the same genome, however, not all the cells look the same nor do they express the same proteins.

One main reason for this difference is that different cells express different genes. Another reason is alternative splicing. Cells can splice RNA transcripts differently to make different proteins from the same gene. In fact, 75% of human genes produce multiple forms of the same proteins which ultimately increases the coding potential of the genome.

Post-transcriptional modifications are also critical for sex determination in the fruit fly. Just like humans, there are certain genes in fruit flies that can be alternatively spliced to form different versions of the same protein: one gene in particular called the doublesex gene is responsible for determining the sex of a fruit fly. Following alternative splicing, one form of the doublesex protein will inhibit genes involved in male fruit fly development.

However, another form of the protein will inhibit female fruit fly development. Importantly, this example illustrates how post-transcriptional modifications can have drastic effects on the organism even though the gene itself can be identical.

Ultimately, alternative splicing increases the coding potential of the genome. Rather than having two independent gene pathways, the same gene can be expressed differently through alternative splicing to have opposite effects.

Doublesex genes determine the sex of a fruit fly by acting as a molecular switch at the end of the signal cascade. These genes guide different types of cells toward male or female differentiation.

Post-Transcriptional Regulation in Prokaryote

Unlike eukaryotes, prokaryotes do not have post-transcriptional modifications. The 5' GTP cap, 3' poly-A tail, and introns are unique to eukaryotes. One reason may be due to the spatial separation between RNA and proteins in eukaryotes.

Unlike eukaryotes, prokaryotes do not have a nucleus so both transcription and translation occur in the cytoplasm. This means that RNA does not need to be transported from the nucleus into the cytoplasm. Without this need for transportation, translation can occur on the mRNA transcript simultaneously as the mRNA transcript is being built.

Therefore, mRNA would not need to be as carefully guarded against degradation since translation starts immediately. Without the need for protection and transport, post-transcriptional modifications would not be needed.

However, prokaryotes have alternative ways to check the quality of the mRNA. If an incomplete or broken mRNA is being translated by the ribosome, an 11 amino acid tag will be added to the end of the newly synthesized polypeptide. This 11 amino tag acts as a signal to proteases within the prokaryotic cell to destroy the entire protein. By destroying the protein, it prevents incomplete or broken proteins from lingering in the cell.

Additionally, without post-transcription modifications, mRNAs are rapidly degraded in the prokaryotic cell; therefore, translation must occur while a new mRNA transcript is being synthesized.

Post-Transcriptional Regulation - Key takeaways

  • Following transcription, a series of enzyme-catalyzed modifications called post-transcriptional modifications occur to convert hnRNA into functional messenger RNA (mRNA).
  • In eukaryotic cells, both transcription and post-transcriptional modifications occur in the nucleus while translation occurs in the cytoplasm.
  • There are three post-transcriptional modifications: addition of a 5' GTP cap, addition of a 3' poly-A tail, and removal of introns.
  • Alternative splicing is the selective inclusion of certain exons so that different forms of a protein can be made from the same transcript.
  • An example of the effects of post-transcriptional modifications is sex determination in the fruit fly.

Frequently Asked Questions about Post-Transcriptional Regulation

Following transcription, a series of enzyme-catalyzed modifications called post-transcriptional modifications occur to convert hnRNA into functional messenger RNA (mRNA).

Following complimentary base pairing of microRNAs (miRNAs) to mRNA, the mRNA transcript will be deactivated and targeted for degradation rather than translation. 

  • Addition of a 5' GTP cap
  • Addition of a 3' poly-A tail
  • Removal of introns and retention of exons 

Lactose digestion is controlled by the lac operon in prokaryotes via the lac operon. The lac operon is regulated before transcription via activator and repressors. 

In eukaryotes, post-transcriptional regulation occurs in the nucleus to hnRNA before being converted to mRNA following export to the cytoplasm. 

Final Post-Transcriptional Regulation Quiz

Post-Transcriptional Regulation Quiz - Teste dein Wissen

Question

The Central Dogma states that ___.

Show answer

Answer

instructions in the form of DNA in the cell are made into RNA before eventually being made into proteins

Show question

Question

During ___ DNA is turned into heterogenous nuclear RNA (hRNA).

Show answer

Answer

transcription

Show question

Question

What are post-transcriptional modifications?

Show answer

Answer

These are enzyme-catalyzed modifications that occur after transcription. These convert hnRNA into functional mRNA.

Show question

Question

In eukaryotic cells, both transcription and post-transcriptional modifications occur in the ___.

Show answer

Answer

nucleus

Show question

Question

Which cells do not have post-transcriptional modifications?

Show answer

Answer

Prokaryotic cells

Show question

Question

Which of the following are the correct post-transcriptional modifications that occur in hnRNA?

Show answer

Answer

Addition of a 5' GTP cap

Show question

Question

What is a 5' GTP cap?

Show answer

Answer

The 5' GTP cap is a modified guanine nucleotide with an additional methyl group at the 7th position of guanine added to the 5' end of hnRNA.

Show question

Question

What are the functions of the 5' GTP cap?

Show answer

Answer

There are three overall functions of the 5' GTP cap. First to aid in RNA processing and export of the RNA from the nucleus. Second, it acts as a marker to orient the mRNA during translation in the cytoplasm. And third, the addition of the 5' GTP cap protects the mRNA from degradation. 

Show question

Question

What is added to the 3' end of the hnRNA?

Show answer

Answer

poly-A tail

Show question

Question

Once any single-stranded RNA strand enters the cytoplasm, it will be rapidly degraded by the cell's degradation machinery known as ____ beginning from its 3' end.

Show answer

Answer

3'-exonucleases

Show question

Question

What is the function of the poly-A tail?

Show answer

Answer

The poly-A tail acts as a buffer that will be degraded before the coding information on the mRNA. The longer the poly-A tail, the longer that the mRNA transcript can survive in the cytoplasm. The poly-A tail is also critical for the export of mature mRNA from the nucleus into the cytoplasm. 

Show question

Question

_____ are non-coding sequences.

Show answer

Answer

Introns

Show question

Question

What is splicing?

Show answer

Answer

The process of removing introns

Show question

Question

Describe how splicesosomes function during splicing.

Show answer

Answer

Splicesosomes remove introns and reassemble exons into one mRNA strand.

Show question

Question

What is alternative splicing?

Show answer

Answer

Alternative splicing is the selective inclusion of exons, allowing for

different forms of the protein to be made from the same DNA sequence. 

Show question

Question

Describe how prokaryotes check the quality of mRNA,

Show answer

Answer

 If an incomplete or broken mRNA is being translated by the ribosome, an 11 amino acid tag will be added to the end of the newly synthesized polypeptide. This 11 amino tag acts as a signal to proteases within the prokaryotic cell to destroy the entire protein. By destroying the protein, it prevents incomplete or broken proteins from lingering in the cell. 

Show question

Test your knowledge with multiple choice flashcards

The Central Dogma states that ___.

During ___ DNA is turned into heterogenous nuclear RNA (hRNA).

In eukaryotic cells, both transcription and post-transcriptional modifications occur in the ___.

Next

Flashcards in Post-Transcriptional Regulation16

Start learning

The Central Dogma states that ___.

instructions in the form of DNA in the cell are made into RNA before eventually being made into proteins

During ___ DNA is turned into heterogenous nuclear RNA (hRNA).

transcription

What are post-transcriptional modifications?

These are enzyme-catalyzed modifications that occur after transcription. These convert hnRNA into functional mRNA.

In eukaryotic cells, both transcription and post-transcriptional modifications occur in the ___.

nucleus

Which cells do not have post-transcriptional modifications?

Prokaryotic cells

Which of the following are the correct post-transcriptional modifications that occur in hnRNA?

Addition of a 5' GTP cap

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