Without mRNA, your genetic code would never get used by your body. Proteins would never get made. Messenger ribonucleic acid, or mRNA for short, plays a vital role in human biology, specifically in a process known as protein synthesis.
At Moderna, we are leveraging the fundamental role that mRNA plays in protein synthesis. We have developed proprietary technologies and methods to create mRNA sequences that cells recognize as if they were produced in the body. Using mRNA as a drug opens up a breadth of opportunities to treat and prevent disease.
Lagging-strand replication is discontinuous, with short Okazaki fragments being formed and later linked together. Molecular biology: Prime-time progress. Nature , All rights reserved. Figure Detail.
One factor that helps ensure precise replication is the double-helical structure of DNA itself. In particular, the two strands of the DNA double helix are made up of combinations of molecules called nucleotides.
DNA is constructed from just four different nucleotides — adenine A , thymine T , cytosine C , and guanine G — each of which is named for the nitrogenous base it contains. Moreover, the nucleotides that form one strand of the DNA double helix always bond with the nucleotides in the other strand according to a pattern known as complementary base-pairing — specifically, A always pairs with T, and C always pairs with G Figure 2.
Thus, during cell division, the paired strands unravel and each strand serves as the template for synthesis of a new complementary strand.
Each nucleotide has an affinity for its partner: A pairs with T, and C pairs with G. In most multicellular organisms, every cell carries the same DNA, but this genetic information is used in varying ways by different types of cells. In other words, what a cell "does" within an organism dictates which of its genes are expressed. Nerve cells, for example, synthesize an abundance of chemicals called neurotransmitters, which they use to send messages to other cells, whereas muscle cells load themselves with the protein-based filaments necessary for muscle contractions.
Transcription is the first step in decoding a cell's genetic information. RNA molecules differ from DNA molecules in several important ways: They are single stranded rather than double stranded; their sugar component is a ribose rather than a deoxyribose; and they include uracil U nucleotides rather than thymine T nucleotides Figure 4.
Also, because they are single strands, RNA molecules don't form helices; rather, they fold into complex structures that are stabilized by internal complementary base-pairing. Messenger RNA mRNA molecules carry the coding sequences for protein synthesis and are called transcripts; ribosomal RNA rRNA molecules form the core of a cell's ribosomes the structures in which protein synthesis takes place ; and transfer RNA tRNA molecules carry amino acids to the ribosomes during protein synthesis. Other types of RNA also exist but are not as well understood, although they appear to play regulatory roles in gene expression and also be involved in protection against invading viruses.
Some mRNA molecules are abundant, numbering in the hundreds or thousands, as is often true of transcripts encoding structural proteins. Other mRNAs are quite rare, with perhaps only a single copy present, as is sometimes the case for transcripts that encode signaling proteins.
In eukaryotes, transcripts for structural proteins may remain intact for over ten hours, whereas transcripts for signaling proteins may be degraded in less than ten minutes.
Cells can be characterized by the spectrum of mRNA molecules present within them; this spectrum is called the transcriptome. Whereas each cell in a multicellular organism carries the same DNA or genome, its transcriptome varies widely according to cell type and function. For instance, the insulin-producing cells of the pancreas contain transcripts for insulin, but bone cells do not.
Even though bone cells carry the gene for insulin, this gene is not transcribed. Therefore, the transcriptome functions as a kind of catalog of all of the genes that are being expressed in a cell at a particular point in time. Figure 5: An electron micrograph of a prokaryote Escherichia coli , showing DNA and ribosomes This Escherichia coli cell has been treated with chemicals and sectioned so its DNA and ribosomes are clearly visible.
The DNA appears as swirls in the center of the cell, and the ribosomes appear as dark particles at the cell periphery. Courtesy of Dr. Abraham Minsky Ribosomes are the sites in a cell in which protein synthesis takes place.
Cells have many ribosomes, and the exact number depends on how active a particular cell is in synthesizing proteins. For example, rapidly growing cells usually have a large number of ribosomes Figure 5.
Ribosomes are complexes of rRNA molecules and proteins, and they can be observed in electron micrographs of cells. Sometimes, ribosomes are visible as clusters, called polyribosomes.
In eukaryotes but not in prokaryotes , some of the ribosomes are attached to internal membranes, where they synthesize the proteins that will later reside in those membranes, or are destined for secretion Figure 6. Although only a few rRNA molecules are present in each ribosome, these molecules make up about half of the ribosomal mass.
The remaining mass consists of a number of proteins — nearly 60 in prokaryotic cells and over 80 in eukaryotic cells. Within the ribosome, the rRNA molecules direct the catalytic steps of protein synthesis — the stitching together of amino acids to make a protein molecule. Eukaryotic and prokaryotic ribosomes are different from each other as a result of divergent evolution. These differences are exploited by antibiotics, which are designed to inhibit the prokaryotic ribosomes of infectious bacteria without affecting eukaryotic ribosomes, thereby not interfering with the cells of the sick host.
Figure 6: The endoplasmic reticulum of this eukaryotic cell is studded with ribosomes. The codons are written 5' to 3', as they appear in the mRNA. Figure Detail But where does translation take place within a cell? What individual substeps are a part of this process? And does translation differ between prokaryotes and eukaryotes?
The answers to questions such as these reveal a great deal about the essential similarities between all species. Within all cells, the translation machinery resides within a specialized organelle called the ribosome. In eukaryotes, mature mRNA molecules must leave the nucleus and travel to the cytoplasm , where the ribosomes are located. On the other hand, in prokaryotic organisms, ribosomes can attach to mRNA while it is still being transcribed.
In all types of cells, the ribosome is composed of two subunits: the large 50S subunit and the small 30S subunit S, for svedberg unit, is a measure of sedimentation velocity and, therefore, mass. Each subunit exists separately in the cytoplasm, but the two join together on the mRNA molecule. The tRNA molecules are adaptor molecules—they have one end that can read the triplet code in the mRNA through complementary base-pairing, and another end that attaches to a specific amino acid Chapeville et al.
The idea that tRNA was an adaptor molecule was first proposed by Francis Crick, co-discoverer of DNA structure, who did much of the key work in deciphering the genetic code Crick, The rRNA catalyzes the attachment of each new amino acid to the growing chain. Interestingly, not all regions of an mRNA molecule correspond to particular amino acids. In particular, there is an area near the 5' end of the molecule that is known as the untranslated region UTR or leader sequence.
This portion of mRNA is located between the first nucleotide that is transcribed and the start codon AUG of the coding region, and it does not affect the sequence of amino acids in a protein Figure 3.
So, what is the purpose of the UTR? It turns out that the leader sequence is important because it contains a ribosome-binding site. A similar site in vertebrates was characterized by Marilyn Kozak and is thus known as the Kozak box. If the leader is long, it may contain regulatory sequences, including binding sites for proteins, that can affect the stability of the mRNA or the efficiency of its translation. Figure 4: The translation initiation complex.
When translation begins, the small subunit of the ribosome and an initiator tRNA molecule assemble on the mRNA transcript. The small subunit of the ribosome has three binding sites: an amino acid site A , a polypeptide site P , and an exit site E.
Here, the initiator tRNA molecule is shown binding after the small ribosomal subunit has assembled on the mRNA; the order in which this occurs is unique to prokaryotic cells.
In eukaryotes, the free initiator tRNA first binds the small ribosomal subunit to form a complex. Figure Detail Although methionine Met is the first amino acid incorporated into any new protein, it is not always the first amino acid in mature proteins—in many proteins, methionine is removed after translation.
In fact, if a large number of proteins are sequenced and compared with their known gene sequences, methionine or formylmethionine occurs at the N-terminus of all of them. However, not all amino acids are equally likely to occur second in the chain, and the second amino acid influences whether the initial methionine is enzymatically removed. For example, many proteins begin with methionine followed by alanine.
In both prokaryotes and eukaryotes, these proteins have the methionine removed, so that alanine becomes the N-terminal amino acid Table 1. However, if the second amino acid is lysine, which is also frequently the case, methionine is not removed at least in the sample proteins that have been studied thus far.
These proteins therefore begin with methionine followed by lysine Flinta et al. Table 1 shows the N-terminal sequences of proteins in prokaryotes and eukaryotes, based on a sample of prokaryotic and eukaryotic proteins Flinta et al.
In the table, M represents methionine, A represents alanine, K represents lysine, S represents serine, and T represents threonine. Once the initiation complex is formed on the mRNA, the large ribosomal subunit binds to this complex, which causes the release of IFs initiation factors. The large subunit of the ribosome has three sites at which tRNA molecules can bind.
The A amino acid site is the location at which the aminoacyl-tRNA anticodon base pairs up with the mRNA codon, ensuring that correct amino acid is added to the growing polypeptide chain. The P polypeptide site is the location at which the amino acid is transferred from its tRNA to the growing polypeptide chain. Finally, the E exit site is the location at which the "empty" tRNA sits before being released back into the cytoplasm to bind another amino acid and repeat the process.
The ribosome is thus ready to bind the second aminoacyl-tRNA at the A site, which will be joined to the initiator methionine by the first peptide bond Figure 5. Figure 5: The large ribosomal subunit binds to the small ribosomal subunit to complete the initiation complex.
The initiator tRNA molecule, carrying the methionine amino acid that will serve as the first amino acid of the polypeptide chain, is bound to the P site on the ribosome.
The A site is aligned with the next codon, which will be bound by the anticodon of the next incoming tRNA. Next, peptide bonds between the now-adjacent first and second amino acids are formed through a peptidyl transferase activity. For many years, it was thought that an enzyme catalyzed this step, but recent evidence indicates that the transferase activity is a catalytic function of rRNA Pierce, After the peptide bond is formed, the ribosome shifts, or translocates, again, thus causing the tRNA to occupy the E site.
The tRNA is then released to the cytoplasm to pick up another amino acid. In addition, the A site is now empty and ready to receive the tRNA for the next codon. This process is repeated until all the codons in the mRNA have been read by tRNA molecules, and the amino acids attached to the tRNAs have been linked together in the growing polypeptide chain in the appropriate order. At this point, translation must be terminated, and the nascent protein must be released from the mRNA and ribosome.
No tRNAs recognize these codons. Thus, in the place of these tRNAs, one of several proteins, called release factors, binds and facilitates release of the mRNA from the ribosome and subsequent dissociation of the ribosome. The translation process is very similar in prokaryotes and eukaryotes. Although different elongation, initiation, and termination factors are used, the genetic code is generally identical. As previously noted, in bacteria, transcription and translation take place simultaneously, and mRNAs are relatively short-lived.
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