RIBOSOMES (PROTEIN FACTORY)

Ribosomes are small, dense, rounded and granular particle of ribonucleo protein. Ribosomes have been found to be the universal component of all biological organisms as they occur in all prokaryotic and eukaryotic cells. They are used by the cell for protein synthesis; the process by which amino acids are assembled in a definite sequence to produce a polypeptide chain. The ribosomes were first noted in plant cells by Robinson and Brown in 1953 when studying bean roots and Palade observed them in animal cells in 1955. He isolated the ribosomes and detected the RNA in them in 1956. Presen knowledge about the ribosomes have been gained by cell fractionation, ultra centrifugation, radioisotope studies and electron microscopy. Occurance and distribution Ribosomes occur in both prokaryotic and eukaryotic cells. The 70S ribosome is made up of a 50S and 30S subunits. The 50S subunit contains the 23S and 5S rRNA while the 30S subunit contains the 16S rRNA. Eukaryotes have 80S ribosomes, each consisting of a small (40S) and large (60S) subunit. Their large subunit is composed of a 5S RNA (120 nucleotides), a 28S RNA (4700 nucleotides), a 5.8S subunit (160 nucleotides) and ~49 proteins. The 40S subunit has a 1900 nucleotide (18S) RNA and ~33 proteins. Eukaryotic ribosome differ from prokaryotic ribosomes in the following respects 1. 2. 3. 4. 5. 6. They are considerably larger. They contain a large no of proteins. (70 80 instead of 53) They have four mplecules of RNA instead of three in prokaryotes. Their proteins and nucleic acids are larger. The RNA- protein ratio is 1:1 instead of 2:1 in prokaryotes. All prokaryotes have 70S (where S=Svedberg units) ribosomes while eukaryotes contain larger 80S ribosomes in their cytosol. 7. Ribosomes occur free forms in prokaryotes and are then called monosomes or may be associated with mRNA to for polysomes. In eukaryotes ribosomes are usually associated with membranes of ER. The 80S ribosome form clusters called polyribosomes or polysomes during protein synthesis.

Types of Ribosomes Ribosome type 80S 80S 70S 55S Found in Eukaryotic animals Eukaryotic plants Prokaryotes Mitochondria (vertibrates) Large subunit 60S 60S 50S 40S 28-29S +5S +5.8S 25S + 5S + 5.8s 23S + 5S 16-17S + 5S Small subunit 40S 40S 30S 30s 18S 16-18S 16S 12-13S

Structure The ribosomes are oblate, spheroid structures of 150-250 in diameter. Each ribosome is porous, hydrated and composed of two subunits. One ribosomal subunit is large in size and has a dome like shape, while the other ribosomal subunit is smaller in size and occurring above the large subunit and forming a cap like structure. Prokaryotes have 70S ribosomes, each consisting of a small (30S) and a large (50S) subunit. Their large subunit is composed of a 5S RNA subunit (consisting of 120 nucleotides), a 23S RNA subunit (2900 nucleotides) and 34 proteins. The 30S subunit has a 1540 nucleotide RNA subunit (16S) bound to 21 proteins. Eukaryotes have 80S ribosomes, each consisting of a small (40S) and large (60S) subunit. Their large subunit is composed of a 5S RNA (120 nucleotides), a 28S RNA (4700 nucleotides), a 5.8S subunit (160 nucleotides) and ~49 proteins. The 40S subunit has a 1900 nucleotide (18S) RNA and ~33 proteins. The ribosomes found in chloroplasts and mitochondria of eukaryotes also consist of large and small subunits bound together with proteins into one 70S particle. These organelles are believed to be descendants of bacteria and as such their ribosomes are similar to those of bacteria. The various ribosomes share a core structure, which is quite similar despite the large differences in size. Much of the RNA is highly organized into various tertiary structural motifs, for example pseudoknots that exhibit coaxial stacking. The extra RNA in the larger ribosomes is in several long continuous insertions, such that they form loops out

of the core structure without disrupting or changing it. All of the catalytic activity of the ribosome is carried out by the RNA; the proteins reside on the surface and seem to stabilize the structure. The two ribosomal subunits remain united with each other due to low concentration of the Mg++ (.001M) ions. When the concentrationof the Mg++ ions reduces in the matrix both ribosomal units separated. In bacterial cells two subunits are found to occur freely in the cytoplasm and they unit only during the process of protein synthesis. At high concentration of the Mg++ ions (0.01M) in th matrix the two ribosomes combined to from a dimer with twice the molecular weight of the individual. Thus Mg++ ions are required by ribosomes for their structural cohesion. During protein synthesis many ribosomes are aggregated due to common mRNA and form the polyribosomes or polysomes FUNCTION Ribosomes are the workhorses of protein biosynthesis, the process of translating mRNA into protein. The mRNA comprises a series of codons that dictate to the ribosome the sequence of the amino acids needed to make the protein. Using the mRNA as a template, the ribosome traverses each codon (3 nucleotides) of the mRNA, pairing it with the appropriate amino acid provided by a tRNA. Molecules of transfer RNA (tRNA) contain a complementary anticodon on one end and the appropriate amino acid on the other. The small ribosomal subunit, typically bound to a tRNA containing the amino acid methionine, binds to an AUG codon on the mRNA and recruits the large ribosomal subunit. The ribosome then contains three RNA binding sites, designated A, P and E. The A site binds an aminoacyl-tRNA (a tRNA bound to an amino acid); the P site binds a peptidyl-tRNA (a tRNA bound to the peptide being synthesized); and the E site binds a free tRNA before it exits the ribosome. Protein synthesis begins at a start codon AUG near the 5' end of the mRNA. mRNA binds to the P site of the ribosome first. The ribosome is able to identify the start codon by use of the Shine-Dalgarno sequence of the mRNA in prokaryotes and Kozak box in eukaryotes.

Translation of mRNA (1) by a ribosome (2)(shown as small and large subunits) into a polypeptide chain (3). The ribosome begins at the start codon of mRNA (AUG) and ends at the stop codon (UAG). In Figure, both ribosomal subunits (small and large) assemble at the start codon (towards the 5' end of the mRNA). The ribosome uses tRNA that matches the current codon (triplet) on the mRNA to append an amino acid to the polypeptide chain. This is done for each triplet on the mRNA, while the ribosome moves towards the 3' end of the mRNA. Usually in bacterial cells, several ribosomes are working parallel on a single mRNA, forming what is called a polyribosome or polysome.