October 15, 2012

Many ribosomes bind to one mRNA

  • A polyribosome (Polysome) is an mRNA that is simultaneously being translated by several ribosomes.
  • A nascent protein has not yet completed its synthesis; the polypeptide chain is still attached to the ribosome via a tRNA.
  • An mRNA is simultaneously translated by several ribosomes. Each ribosome is at a different stage of progression along the mRNA. 

When active ribosomes are isolated in the form of the fraction associated with newly synthesized proteins, they are found in the form of a complex consisting of an mRNA associated with several ribosomes. This is the polyribosome or polysome. The 30S subunit of each ribosome is associated with the mRNA, and the 50S subunit carries the newly synthesized protein. The tRNA spans both subunits.

Each ribosome in the polysome independently synthesizes a single polypeptide during its traverse of the messenger sequence. Essentially the mRNA is pulled through the ribosome, and each triplet nucleotide is translated into an amino acid. So the mRNA has a series of ribosomes that carry increasing lengths of the protein product, moving from the 5 to the 3 end, as illustrated in Figure 5.9. A polypeptide chain in the process of synthesis is sometimes called a nascent protein.
Roughly the most recent 30-35 amino acids added to a growing polypeptide chain are protected from the environment by the structure of the ribosome. Probably all of the preceding part of the polypeptide protrudes and is free to start folding into its proper conformation. So proteins can display parts of the mature conformation even before synthesis has been completed.  

A classic characterization of polysomes is shown in the electron micrograph of Figure 5.10. Globin protein is synthesized by a set of 5 ribosomes attached to each mRNA (pentasomes). The ribosomes appear as squashed spherical objects of ~7 nm (70 Å) in diameter, connected by a thread of mRNA. The ribosomes are located at various positions along the messenger. Those at one end have just started protein synthesis; those at the other end are about to complete production of a polypeptide chain (Slayter et al., 1963).
The size of the polysome depends on several variables. In bacteria, it is very large, with tens of ribosomes simultaneously engaged in translation. Partly the size is due to the length of the mRNA (which usually codes for several proteins); partly it is due to the high efficiency with which the ribosomes attach to the mRNA.
Polysomes in the cytoplasm of a eukaryotic cell are likely to be smaller than those in bacteria; again, their size is a function both of the length of the mRNA (usually representing only a single protein in eukaryotes) and of the characteristic frequency with which ribosomes attach. An average eukaryotic mRNA probably has ~8 ribosomes attached at any one time.

Figure 5.11 illustrates the life cycle of the ribosome. Ribosomes are drawn from a pool (actually the pool consists of ribosomal subunits), used to translate an mRNA, and then return to the pool for further cycles. The number of ribosomes on each mRNA molecule synthesizing a particular protein is not precisely determined, in either bacteria or eukaryotes, but is a matter of statistical fluctuation, determined by the variables of mRNA size and efficiency. 

An overall view of the attention devoted to protein synthesis in the intact bacterium is given in Figure 5.12. The 20,000 or so ribosomes account for a quarter of the cell mass. There are >3000 copies of each tRNA, and altogether, the tRNA molecules outnumber the ribosomes by almost tenfold; most of them are present as aminoacyl-tRNAs, that is, ready to be used at once in protein synthesis. Because of their instability, it is difficult to calculate the number of mRNA molecules, but a reasonable guess would be ~1500, in varying states of synthesis and decomposition. There are ~600 different types of mRNA in a bacterium. This suggests that there are usually only 2-3 copies of each mRNA per bacterium. On average, each probably codes for ~3 proteins. If there are 1850 different soluble proteins, there must on average be >1000 copies of each protein in a bacterium.