Ribosome Structure and Function


 

Ribosomes are cytoplasmic organelles found in prokaryotes and eukaryotes. They are large complexes of proteins and three (prokaryotes) or four (eukaryotes) rRNA (ribosomal ribonucleic acid) molecules called subunits made in the nucleolus. The main function of ribosomes is to serve as the site of mRNA translation ( = protein synthesis, the assembly of amino acids into proteins); once the two (large and small) subunits are joined by the mRNA from the nucleus, the ribosome translates the mRNA into a specific sequence of amino acids, or a polypeptide chain.

 

Ribosome

 

Type and Location of Ribosomes

Ribosomes are found in cells in two ways: free and bound. In an electron micrograph as shown above, ribosomes appear as dark granules. Ribosomes exist at various locations within the cell; however, the location of ribosomes depends on the function of the cell:

Free ribosomes

Bound ribosomes

Ribosomes are also located in mitochondria and chloroplasts of eukaryotic cells; they are always smaller than cytoplasmic ribosomes and are comparable to prokaryotic ribosomes in both size and sensitivity to anitbiotics; however, the sedimentation values s (s = Svedberg units: a measure of the rate of sedimentation of a component in a centrifuge, related both to the molecular weight and the 3-D shape of the component) vary somewhat in different phyla. Prokaryotic and eukaryotic ribosomes perform the same functions by the same set of chemical reactions; however, eukaryotic ribosomes are much larger than prokaryotic ones and most of their proteins are different. Mitochondrial and chloroplast ribosomes resemble bacterial ribosomes.

Cells put forth considerable effort to the production of these essential organelles; for example, one E. coli cell contains about 15,000 ribosomes, each one with a molecular weight of approximately 3 x 106 daltons constituting 25% of the total mass of these bacterial cells.

 

Structure of Ribosomes

Subunits

Form: Small and Large Subunits

Prokaryotic and eukaryotic ribosomes are very similar in their form. Small prokaryotic and eukaryotic ribosomal subunits have a head and a base with an armlike platform extending from one side as seen below left; however, additional features of the small eukaryotic ribosome subunit include a bill that extends from the head of the small subunit on the side opposite the cleft and a set of lobes at the end of the subunit opposite the head (below right); the lobes are believed to contain the additional sequences that make 18s rRNA larger that its 16s bacterial counterpart.

Prokaryote Small

The large subunit has a prominent central protuberance, stalk, and ridge extending from one side as seen pictured above right. The large subunit has a tunnel about 10nm long and 2.5 nm in diameter; the tunnel extends from the region containing the A (aminoacyl) and P (peptidyl) sites to the part of the large subunit from which the newly assembled polypeptide chain exits the ribosome. This tunnel is thought to be the channel that newly assembled polypeptide chains travel on the way out of the ribosome.

Components

Arrangement

Ribosomes are small, but complex structures, roughly 20 to 30 nm in diameter, consisting of two unequally sized subunits, referred to as large and small which fit closely together as seen below. A subunit is composed of a complex between RNA molecules and proteins; each subunit contains at least one ribosomal RNA (rRNA) subunit and a large quantity of ribosomal proteins. The subunits together contain up to 82 specific proteins assembled in a precise sequence.

 

Pro & Euk Struct

The prokaryotic ribosome in E. coli has a size of 70s. The two subunits have distinct and recognizable 3-D shapes. Approximately two-thirds of the E. coli ribosome consists of rRNA with the rest consisting of ribosomal proteins. Overall, the 50s and 30s subunits combined are 70s due to the 3-D shape of the ribosomes.

The Components of the Prokaryotic Ribsome

Type of rRNA
Approximate number of nucleotides
Subunit Location
16s
1,542
30s
5s
120
50s
23s
2,904
50s

 

In general, eukaryotic ribosomes are larger and more complex than prokaryotic ribosomes. The size of the ribosome and the molecular weights of the rRNA molecules differ from organism to organism. Simple eukaryotes have the smallest ribosomes, although they are larger than E. coli ribosomes, while mammals have the largest ribosomes. Nonetheless, all eukaryotic ribosomes have many common structural and chemical features. For example, the mammalian ribosome has a size of 80s consisting of a large 60s subunit and a small 40s subunit. The 80s mammalian ribosome consists of about equal weights of rRNA and ribosomal proteins.

 

The Components of the Eukaryotic Ribosome

Type of rRNA
Approximate number of nucleotides
Subunit Location
18s
1,900
40s
5s
120
60s
5.8s
156
60s
28s
4,700
60s

 

Function of Ribosomes

The nucleus is the ultimate control center for cell activities, including translation. Within the chromatin, information required for cellular protein synthesis is coded into the DNA; each DNA segment containing the information for making a protein consitiutes a gene. The information in a protein-encoding gene is copied into a messenger RNA (mRNA) molecule that moves to the cytoplasm through the pores of the nuclear envelope. In the cytoplasm, mRNA molecules are used by ribosomes as directions for protein assembly. The DNA of the entire nucleus contains the codes for many thousands of different proteins.

Pro & Euk Cells

In protein synthesis, a ribosome moves along an mRNA molecule, reading the codon for protein assembly as it goes. As it moves, the ribosome assembles amino acids into a guradually lengthening protein chain. At the end of the coded message, translation stops, the ribosomal subunits separate and detach from the mRNA, and the completed protein is realased. Transfer RNA (tRNA) molecules function as the "dictionary" in the translation mechanism. Each of the 20 amino acids used in protein synthesis is linked to a specific kind of tRNA. The tRNA is capable is recognizing and binding the nucleid acid code word (called a codon) specifiying its attached amino acid in an mRNA molecule.

Overview of Translation

1. Formation of the initiation complex

2. Elongation of the polypeptide chain (one repetition of the steps
a, b and c for every amino acid incoporated into the protein being synthesized:
a. binding of the aminoacyl-tRNA
b. peptide bond formation
c. tranlocation
3. Termination

Steps in Translation

  1. Components necessary for the initiation of translation include the interaction of the small and large ribosomal subunits, mRNA, an initiator aminoacyl-tRNA, GTP, and a large group of initiation factors. The initiation complex is formed by the small ribosomal subunit, the mRNA and Met-tRNA (methionine attached to a specialized initiator tRNA type). The Met-tRNA is hydrogen bonded to the AUG initiation codon on the mRNA
  2. Formation of the ribosomal initiation complex is completed with the addition of the large ribosomal subunit. The AUG codon and the Met-tRNA are positioned in the P (peptidyl) site of the ribosome.
  3. New aminoacyl-tRNA is then positioned in the A (aminoacyl) site.
  4. The covalent bond between the amino acid and the tRNA in the P site is broken.
  5. A peptide (covalent) bond is formed bewteen the two amino acids.
  6. The empty tRNA then dissociates from the P site.
  7. Translocation of the ribosome occurs such that the peptidyl-tRNA in the A site is translocated into the P site.
  8. The complex now looks very similar to that at the initiation of tranlstion. The peptidyl-tRNA is in the P site and the A site is empty and ready to accept the next aminoacyl-tRNA.
  9. The amino acid in the P site is separated from its tRNA and peptide bond formation takes place with the aminoacyl-tRNA in the A site. The tRNA is liberated from the P site and the ribosome tranlocates such that the new peptidyl-tRNA is in the P site; the A site is ready to accept the next aminoacyl-tRNA.
  10. The elogation cycle (the addition of amino acids one at a time to a growing polypeptide chain) continues for more peptide bond formation and translocation.
  11. A stop codon (UAA, UAG, or UGA) positions in the A site; a stop codon has no anticodon (an aminoacyl tRNA that will hydrogen bond to this codon); for this reason, no other amino acid will be added.
  12. The stop codon is regocnized and bound by a protein called the termination or release factor.
  13. The release factor binds to the stop codon at the A site and begins a sequence of events that brings about the termination of translation.
  14. The termination sequence begin with the dissociation of the newly synthesized protein from the peptidyl-tRNA in the P site.
  15. Separation of the newly synthesized proteins from the ribosome is followed by the dissocation of all the remaining subunits.
  16. The ribosomal subunits and the mRNA can reassemble with Met-tRNA to form new initation complexes and prteins translation can begin again to produce additional copies of the protein.

Initiation

  1. A ribosome separates into large and small subunits
  2. Met-tRNA combines with GTP in a side reaction involving an initiation factor
  3. Met-tRNA is added to the small ribosomal subunit
  4. The small subunit is added to the mRNA in a reaction driven by ATP hydrolysis; attachment takes place at the 5' cap of the mRNA; once attached, tha small subunits moves or "scans" along the mRNA until it reaches the AUG initiator codon
  5. The large ribosomal subunit is added driven by the hydrolysis of GTP brought to the complex with the initiator tRNA; elongation follows

Elongation

  1. Aminoacyl-tRNA binds to the A site
  2. Peptide bond formation
  3. Peptidyl-tRNA formed at the A site by step two is transferred from the A site to the P site

 

Peptide Bond Formation

Peptide bond formation is catalyzed on the ribosome by peptidyl transferase. (a) Adjacent aminoacyl-tRNAs bound to the mRNA at the ribosome (b) folowing peptide bond formation, an uncharged tRNA is in the P site and a dipeptidyl-tRNA in the A site.

 

Termination of Translation

The ribosome recognizes a chain termination codon (here, UAG) with the aid of release factors. The release factor reads the stop codon, and this initiatiartes a series of specific termination events leading to the release of the completed polypeptide

  1. Stop codon is encountered at the A site which causes the release factor to bind to the A site along with GTP insead of aminoacyl-tRNA
  2. The release factor binds to the stop codon and hydrolysis of the bond holding the polypeptide chain to the tRNA site at the P site, catalyzed by the peptidyl tranferase site of the large subunit
  3. Since there is no amino acid located at the A site, the hydrolysis allows the polypeptide chain to be freed from the ribosome; with the release of the polypeptide, the release factor is ejected from the A site, and the empty tRNA is ejected from the P site
  4. Ribosomal components separpate










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