In this article, we define RNA polymerase and further explore its various functions throughout cell biology. RNA polymerase in action. While prokaryotic RNA polymerase can directly bind to DNA promoter sequences, eukaryotic forms require the assistance of transcription factors for initial binding. Once RNA polymerase successfully binds DNA at the targeted promoter region, the enzyme can continue with the second stage of transcription.
Termination is the final step of transcription. Once RNA polymerase encounters a terminator sequence or signal, it stops adding complementary nucleotides to the RNA strand. What are the different types of RNA polymerase? While prokaryotes like bacteria have one RNA polymerase that transcribes all types of RNA, eukaryotes like plants and mammals can have numerous forms of RNA polymerase. These transcripts are produced within the nucleolus, a region within the nucleus where ribosomes are assembled.
The availability of rRNA molecules produced by RNA polymerase can impact essential functions of cell biology since these transcripts are directly involved with the production of ribosomes. This subunit enzyme works as a complex that directly influences gene expression through its production of pre-mRNA transcripts. Transcription takes place in the nucleus.
Figure 1 shows how this occurs. Figure 1. Overview of Transcription. Triplets are groups of three successive nucleotide bases in DNA. Codons are complementary groups of bases in mRNA. 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. This page has been archived and is no longer updated.
DNA safely and stably stores genetic material in the nuclei of cells as a reference, or template. Meanwhile, mRNA is comparable to a copy from a reference book because it carries the same information as DNA but is not used for long-term storage and can freely exit the nucleus.
Transcription is carried out by an enzyme called RNA polymerase and a number of accessory proteins called transcription factors.
0コメント