The Ribosome assembles polymeric protein molecules whose sequence is controlled by the sequence messenger RNA molecule and which is required by all living cells or associated viruses).

The ribosome is a large complex molecule which is responsible for catalyzing the formation of proteins from individual amino acids using messenger RNA as a template.^[1] This process is known as translation. Ribosomes are found in all living cells.

The sequence of DNA encoding for a protein may be copied many times into messenger RNA (mRNA) chains of a similar sequence. Ribosomes can bind to an mRNA chain and use it as a template for determining the correct sequence of amino acids in a particular protein. Amino acids are selected, collected and carried to the ribosome by transfer RNA (tRNA molecules), which enter one part of the ribosome and bind to the messenger RNA chain. The attached amino acids are then linked together by another part of the ribosome.

More than one ribosome may move along a single mRNA chain at one time, each "reading" its sequence and producing a corresponding protein molecule. Once the protein is produced it can then 'fold up' to produce a specific functional three-dimensional structure largely determined probabilistically by the pattern of charges in its sequence.

A ribosome is made from complexes of RNAs and proteins called ribonucleoproteins. Each ribosome is divided into two subunits. The smaller subunit binds to the mRNA pattern, while the larger subunit binds to the tRNA and the amino acids. When a ribosome finishes reading an mRNA molecule, these two subunits split apart. Ribosomes have been classified as ribozymes, because the ribosomal RNA seems to be most important for the peptidyl transferase activity that links amino acids together.

Ribosomes from bacteria, archaea and eukaryotes (the three domains of life on Earth), have significantly different structures and RNA sequences. These differences in structure allow some antibiotics to kill bacteria by inhibiting their ribosomes, while leaving human ribosomes unaffected. The ribosomes in the mitochondria of eukaryotic cells functionally resemble in many features those in bacteria, reflecting the likely evolutionary origin of mitochondria.^[2][3] The word ribosome comes from ribo nucleic acid and the Greek: soma (meaning body).

Together with Albert Claude and Christian de Duve, George Emil Palade was awarded the Nobel Prize in Physiology or Medicine, in 1974, for the discovery of the ribosomes.^[4] The Nobel Prize in Chemistry 2009 was awarded to Venkatraman Ramakrishnan, Thomas A. Steitz and Ada E. Yonath for discovering the detailed structure and mechanism of the ribosome.^[5]

Description[link]

Archaeal, eubacterial and eukaryotic ribosomes differ in their size, composition and the ratio of protein to RNA. Because they are formed from two subunits of non-equal size, they are slightly longer in the axis than in diameter. Prokaryotic ribosomes are around 20 nm (200 Å) in diameter and are composed of 65% ribosomal RNA and 35% ribosomal proteins. Eukaryotic ribosomes are between 25 and 30 nm (250–300 Å) in diameter and the ratio of rRNA to protein is close to 1. Ribosomes translate messenger RNA (mRNA) and build polypeptide chains (e.g., proteins) using amino acids delivered by transfer RNA (tRNA). Their active sites are made of RNA, so ribosomes are now classified as "ribozymes".^[6]

Ribosomes build proteins from the genetic instructions held within messenger RNA. Free ribosomes are suspended in the cytosol (the semi-fluid portion of the cytoplasm); others are bound to the rough endoplasmic reticulum, giving it the appearance of roughness and thus its name, or to the nuclear envelope. As ribozymes are partly constituted from RNA, it is thought that they might be remnants of the RNA world.^[7] Although catalysis of the peptide bond involves the C2 hydroxyl of RNA's P-site (see Function section below) adenosine in a protein shuttle mechanism, other steps in protein synthesis (such as translocation) are caused by changes in protein conformations.

Ribosomes are sometimes referred to as organelles, but the use of the term organelle is often restricted to describing sub-cellular components that include a phospholipid membrane, which ribosomes, being entirely particulate, do not. For this reason, ribosomes may sometimes be described as "non-membranous organelles".

Ribosomes were first observed in the mid-1950s by Romanian cell biologist George Emil Palade using an electron microscope as dense particles or granules^[8] for which, in 1974, he would win a Nobel Prize. The term "ribosome" was proposed by scientist Richard B. Roberts in 1958:

During the course of the symposium a semantic difficulty became apparent. To some of the participants, "microsomes" mean the ribonucleoprotein particles of the microsome fraction contaminated by other protein and lipid material; to others, the microsomes consist of protein and lipid contaminated by particles. The phrase “microsomal particles” does not seem adequate, and “ribonucleoprotein particles of the microsome fraction” is much too awkward. During the meeting, the word "ribosome" was suggested, which has a very satisfactory name and a pleasant sound. The present confusion would be eliminated if “ribosome” were adopted to designate ribonucleoprotein particles in sizes ranging from 35 to 100S.

— Roberts, R. B., Microsomal Particles and Protein Synthesis^[9]

The structure and function of the ribosomes and associated molecules, known as the translational apparatus, has been of research interest since the mid-twentieth century and is a very active field of study today.

Figure 2 : Large (red) and small (blue) subunit fit together

Ribosomes consist of two subunits (Figure 1) that fit together (Figure 2) and work as one to translate the mRNA into a polypeptide chain during protein synthesis (Figure 3). Bacterial subunits consist of one or two and eukaryotic of one or three very large RNA molecules (known as ribosomal RNA or rRNA) and multiple smaller protein molecules. Crystallographic work has shown that there are no ribosomal proteins close to the reaction site for polypeptide synthesis. This suggests that the protein components of ribosomes act as a scaffold that may enhance the ability of rRNA to synthesize protein rather than directly participating in catalysis (See: Ribozyme).

Biogenesis[link]

In bacterial cells, ribosomes are synthesized in the cytoplasm through the transcription of multiple ribosome gene operons. In eukaryotes, the process takes place both in the cell cytoplasm and in the nucleolus, which is a region within the cell nucleus. The assembly process involves the coordinated function of over 200 proteins in the synthesis and processing of the four rRNAs, as well as assembly of those rRNAs with the ribosomal proteins.

Ribosome locations[link]

Ribosomes are classified as being either "free" or "membrane-bound".

A ribosome translating a protein that is secreted into the endoplasmic reticulum.

Free and membrane-bound ribosomes differ only in their spatial distribution; they are identical in structure. Whether the ribosome exists in a free or membrane-bound state depends on the presence of an ER-targeting signal sequence on the protein being synthesized, so an individual ribosome might be membrane-bound when it is making one protein, but free in the cytosol when it makes another protein.

Free ribosomes[link]

Free ribosomes can move about anywhere in the cytosol, but are excluded from the cell nucleus and other organelles. Proteins that are formed from free ribosomes are released into the cytosol and used within the cell. Since the cytosol contains high concentrations of glutathione and is, therefore, a reducing environment, proteins containing disulfide bonds, which are formed from oxidized cysteine residues, cannot be produced in this compartment.

Membrane-bound ribosomes[link]

When a ribosome begins to synthesize proteins that are needed in some organelles, the ribosome making this protein can become "membrane-bound". In eukaryotic cells this happens in a region of the endoplasmic reticulum (ER) called the "rough ER". The newly produced polypeptide chains are inserted directly into the ER by the ribosome undertaking vectorial synthesis and are then transported to their destinations, through the secretory pathway. Bound ribosomes usually produce proteins that are used within the plasma membrane or are expelled from the cell via exocytosis.^[10]

Structure[link]

Atomic structure of the 30S Subunit from Thermus thermophilus. Proteins are shown in blue and the single RNA strand in orange. It is found by MRC Laboratory of Molecular Biology in Cambridge, England.^[11]

The ribosomal subunits of prokaryotes and eukaryotes are quite similar.^[12]

The unit of measurement is the Svedberg unit, a measure of the rate of sedimentation in centrifugation rather than size and accounts for why fragment names do not add up (70S is made of 50S and 30S).

Prokaryotes have 70S ribosomes, each consisting of a small (30S) and a large (50S) subunit. Their small subunit has a 16S RNA subunit (consisting of 1540 nucleotides) bound to 21 proteins. The large subunit is composed of a 5S RNA subunit (120 nucleotides), a 23S RNA subunit (2900 nucleotides) and 31 proteins.^[12] Affinity label for the tRNA binding sites on the E. coli ribosome allowed the identification of A and P site proteins most likely associated with the peptidyltransferase activity; labelled proteins are L27, L14, L15, L16, L2; at least L27 is located at the donor site, as shown by E. Collatz and A.P. Czernilofsky.^[13][14] Additional research has demonstrated that the S1 and S21 proteins, in association with the 3'-end of 16S ribosomal RNA, are involved in the initiation of translation.^[15]

Eukaryotes have 80S ribosomes, each consisting of a small (40S) and large (60S) subunit. Their 40S subunit has an 18S RNA (1900 nucleotides) and 33 proteins.^[16] The large subunit is composed of a 5S RNA (120 nucleotides), 28S RNA (4700 nucleotides), a 5.8S RNA (160 nucleotides) subunits and ~49 proteins.^[12][17] During 1977, Czernilofsky published research that used affinity labeling to identify tRNA-binding sites on rat liver ribosomes. Several proteins, including L32/33, L36, L21, L23, L28/29 and L13 were implicated as being at or near the peptidyl transferase center.^[18]

The ribosomes found in chloroplasts and mitochondria of eukaryotes also consist of large and small subunits bound together with proteins into one 70S particle.^[12] These organelles are believed to be descendants of bacteria (see Endosymbiotic theory) and as such their ribosomes are similar to those of bacteria.^[19]

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.^[12] 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.^[12]

The differences between the bacterial and eukaryotic ribosomes are exploited by pharmaceutical chemists to create antibiotics that can destroy a bacterial infection without harming the cells of the infected person. Due to the differences in their structures, the bacterial 70S ribosomes are vulnerable to these antibiotics while the eukaryotic 80S ribosomes are not.^[20] Even though mitochondria possess ribosomes similar to the bacterial ones, mitochondria are not affected by these antibiotics because they are surrounded by a double membrane that does not easily admit these antibiotics into the organelle.^[21]

High-resolution structure[link]

Atomic structure of the 50S Subunit from Haloarcula marismortui. Proteins are shown in blue and the two RNA strands in orange and yellow.^[22] The small patch of green in the center of the subunit is the active site.

The general molecular structure of the ribosome has been known since the early 1970s. In the early 2000s the structure has been achieved at high resolutions, on the order of a few Å.

The first papers giving the structure of the ribosome at atomic resolution were published in rapid succession in late 2000. First, the 50S (large prokaryotic) subunit from the archaeon Haloarcula marismortui was published.^[22] Soon after, the structure of the 30S subunit from Thermus thermophilus was published.^[11] Shortly thereafter, a more detailed structure was published.^[23] These structural studies were awarded the Nobel Prize in Chemistry in 2009. Early the next year (May 2001) these coordinates were used to reconstruct the entire T. thermophilus 70S particle at 5.5 Å resolution.^[24]

Two papers were published in November 2005 with structures of the Escherichia coli 70S ribosome. The structures of a vacant ribosome were determined at 3.5-Å resolution using x-ray crystallography.^[25] Then, two weeks later, a structure based on cryo-electron microscopy was published,^[26] which depicts the ribosome at 11–15 Å resolution in the act of passing a newly synthesized protein strand into the protein-conducting channel.

The first atomic structures of the ribosome complexed with tRNA and mRNA molecules were solved by using X-ray crystallography by two groups independently, at 2.8 Å^[27] and at 3.7 Å.^[28] These structures allow one to see the details of interactions of the Thermus thermophilus ribosome with mRNA and with tRNAs bound at classical ribosomal sites. Interactions of the ribosome with long mRNAs containing Shine-Dalgarno sequences were visualized soon after that at 4.5- to 5.5-Å resolution.^[29]

More recently, in 2010 cryoelectron microscopy was used in determining the first complete atomic model of a eukaryotic 40S ribosomal structure in Tetrahymena thermophila. This research has revealed the structure of the 18S ribosomal RNA and all ribosomal proteins of the 40S subunit, as well as much about the 40S subunit's interaction with eIF1 during translation initiation.^[16] Similarly, in 2011 the eukaryotic 60S subunit structure was also determined from Tetrahymena thermophila in complex with eIF6.^[17]

Function[link]

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.

Figure 3 : 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 3, 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.

See also[link]

References[link]

External links[link]

Wikimedia Commons has media related to: Ribosomes

• 70S Ribosome Architecture Animation of a working ribosome. Requires the Chime browser plugin from this site (where registration is required).
• Lab computer simulates ribosome in motion
• Role of the Ribosome, Gwen V. Childs, copied here
• Ribosome in Proteopedia - The free, collaborative 3D encyclopedia of proteins & other molecules
• Ribosomal proteins families in ExPASy
• Molecule of the Month © RCSB Protein Data Bank:
  • Ribosome
  • Elongation Factors
  • Palade

 This article incorporates public domain material from the NCBI document "Science Primer".

Ada E. Yonath
Born	(1939-06-22) 22 June 1939 (age 73) Jerusalem
Residence	Israel
Nationality	Israeli
Fields	Crystallography
Institutions	Weizmann Institute of Science
Alma mater	Hebrew University of Jerusalem Weizmann Institute of Science
Known for	Cryo bio-crystallography
Notable awards	Harvey Prize (2002) Wolf Prize in Chemistry (2006) L'Oréal-UNESCO Award for Women in Science (2008) Nobel Prize in Chemistry (2009).

Ada E. Yonath (Hebrew: עדה יונת‎, pronounced [ˈada joˈnat]) (born 22 June 1939^[1]) is an Israeli crystallographer best known for her pioneering work on the structure of the ribosome. She is the current director of the Helen and Milton A. Kimmelman Center for Biomolecular Structure and Assembly of the Weizmann Institute of Science. In 2009, she received the Nobel Prize in Chemistry along with Venkatraman Ramakrishnan and Thomas A. Steitz for her studies on the structure and function of the ribosome, becoming the first Israeli woman to win the Nobel Prize out of ten Israeli Nobel laureates,^[2] the first woman from the Middle East to win a Nobel prize in the sciences,^{[citation needed]} and the first woman in 45 years to win the Nobel Prize for Chemistry. However, she said herself that there was nothing special about a woman winning the Prize.^[3]

Biography[link]

Yonath (née Lifshitz)^[4] was born in the Geula quarter of Jerusalem.^[5] Her parents, Hillel and Esther Lifshitz, were Zionist Jews who immigrated to Palestine from Łódź (Poland) in 1933 before the establishment of Israel.^[6] Her father was a rabbi and came from a rabbinical family. They settled in Jerusalem and ran a grocery, but found it difficult to make ends meet. They lived in cramped quarters with several other families, and Yonath remembers "books" being the only thing she had to keep her occupied.^[7] Despite their poverty, her parents sent her to school in the upscale Beit HaKerem neighborhood to assure her a good education. When her father died at the age of 42, the family moved to Tel Aviv.^[8] Yonath was accepted to Tichon Hadash high school although her mother could not pay the tuition. She gave math lessons to students in return.^[9] As a youngster, she says she was inspired by the Polish-French scientist Marie Curie.^[10] However, she stresses that Curie, whom she as a child was fascinated by after reading a well-written biography, was not her "role model".^[11] She returned to Jerusalem for college, graduating from the Hebrew University of Jerusalem with a bachelor's degree in chemistry in 1962, and a master's degree in biochemistry in 1964. In 1968, she earned a Ph.D. in X-Ray crystallography at the Weizmann Institute of Science.

She has one daughter, Hagit Yonath, a doctor at Sheba Medical Center, and a granddaughter, Noa.^[12] She is the cousin of anti-occupation activist Dr Ruchama Marton.^[13]

She has called for the unconditional release of all Hamas prisoners, saying that "holding Palestinians captive encourages and perpetuates their motivation to harm Israel and its citizens ... once we don't have any prisoners to release they will have no reason to kidnap soldiers".^[14]

Scientific career[link]

Ada Yonath at the Weizmann Institute of Science

Yonath accepted postdoctoral positions at the Carnegie Mellon University (1969) and MIT (1970). While a postdoc at MIT she spent some time in the lab of subsequent 1976 chemistry Nobel Prize winner William N. Lipscomb, Jr. of Harvard University where she was inspired to pursue very large structures.^[15]

In 1970, she established what was for nearly a decade the only protein crystallography laboratory in Israel. Then, from 1979 to 1984 she was a group leader with Heinz-Günter Wittmann at the Max Planck Institute for Molecular Genetics in Berlin. She was visiting professor at the University of Chicago in 1977-78.^[16] She headed a Max-Planck Institute Research Unit at DESY in Hamburg, Germany (1986–2004) in parallel to her research activities at the Weizmann Institute.

Yonath focuses on the mechanisms underlying protein biosynthesis, by ribosomal crystallography, a research line she pioneered over twenty years ago despite considerable skepticism of the international scientific community.^[17] Ribosomes translate RNA into protein and because they have slightly different structures in microbes, when compared to eukaryotes, such as human cells, they are often a target for antibiotics. She determined the complete high-resolution structures of both ribosomal subunits and discovered within the otherwise asymmetric ribosome, the universal symmetrical region that provides the framework and navigates the process of polypeptide polymerization. Consequently she showed that the ribosome is a ribozyme that places its substrates in stereochemistry suitable for peptide bond formation and for substrate-mediated catalysis. Two decades ago she visualized the path taken by the nascent proteins, namely the ribosomal tunnel, and recently revealed the dynamics elements enabling its involvement in elongation arrest, gating, intra-cellular regulation and nascent chain trafficking into their folding space.

Additionally, Yonath elucidated the modes of action of over twenty different antibiotics targeting the ribosome, illuminated mechanisms of drug resistance and synergism, deciphered the structural basis for antibiotic selectivity and showed how it plays a key role in clinical usefulness and therapeutic effectiveness, thus paving the way for structure-based drug design.

For enabling ribosomal crystallography Yonath introduced a novel technique, cryo bio-crystallography, which became routine in structural biology and allowed intricate projects otherwise considered formidable.^[18]

At the Weizmann Institute, Yonath is the incumbent of the Martin S. and Helen Kimmel Professorial Chair.

Awards and honors[link]

Telephone interview with Ada Yonath during the announcement of the Nobel Prize

Yonath is a member of the United States National Academy of Sciences; the American Academy of Arts and Sciences; the Israel Academy of Sciences and Humanities; the European Academy of Sciences and Art and the European Molecular Biology Organization.

Her awards and honors include the following:

• In 2000, the first European Crystallography Prize;
• In 2002, the Israel Prize, for chemistry;^[1][19]
• In 2006, the Wolf Prize in Chemistry (co-recipient with George Feher) "for ingenious structural discoveries of the ribosomal machinery of peptide-bond formation and the light-driven primary processes in photosynthesis";^[20]
• In 2007, the Paul Ehrlich and Ludwig Darmstaedter Prize;
• In 2008, she became the first Israeli woman to win the L'Oréal-UNESCO Award for Women in Science for her vital work identifying how bacteria become resistant to antibiotics;^[8]
• In 2009, the Nobel Prize in Chemistry (co-recipient with Thomas Steitz and Venkatraman Ramakrishnan).^[21] She was the first Israeli woman to be awarded a Nobel Prize;^[22]
• As well as the Harvey Prize from Technion - Israel Institute of Technology, the Kilby Prize, the Cotton Medal of the US Chemical Society, the Anfinsen Award of the International Protein Society, the Paul Karrer Gold Medal from the University of Zurich, the University of Southern California's Massry Award and Medal, the Datta Medal of the Federation of European Biochemical Societies, the Fritz Lipmann Award of the German Biochemical Society and the Louisa Gross Horwitz Prize from Columbia University.

Quotes[link]

"Survival is far more complicated, much more demanding (than doing science)," she says. "You can always try another approach; even change your subject when a scientific strategy or experiment fails. But when you are hungry you are hungry!" (She was exposed to extreme poverty when her father died prematurely. By 11 she was already working and taking care of her family.) ^[23]

See also[link]

• History of RNA biology
• List of Israel Prize recipients
• List of female Nobel laureates
• List of Jewish Nobel laureates
• List of RNA biologists

References[link]

External links[link]

Wikimedia Commons has media related to: Ada E. Yonath

• Information and Resources, from the Office of Scientific and Technical Information, United States Department of Energy
• "APS user shares the “Israeli Nobel” for chemistry", from the Argonne National Laboratory Advanced Photon Source (APS), United States Department of Energy
• The Official Site of Louisa Gross Horwitz Prize
• Weizmann Institute of Science, Yonath-Site
• Talk of Ada Yonath at the Origins 2011 congress