Chapter 27

Which one of the following statements about the elongation phase of protein synthesis is true? A) At least five high-energy phosphoryl groups are expended for each peptide bond formed. B) During elongation, incoming aminoacylated tRNAs are first bound in the P site. C) Elongation factor EF-Tu facilitates translocation. D) Peptidyl transferase catalyzes the attack of the carboxyl group of the incoming amino acid on an ester linkage in the nascent polypeptide. E) Peptidyl transferase is a ribozyme.

E) Peptidyl transferase is a ribozyme.

Which of the following statements about the tRNA that normally accepts phenylalanine is false? (mRNA codons for phenylalanine are UUU and UUC.) A) It interacts specificially with the Phe synthetase. B) It will accept only the amino acid phenylalanine. C) Its molecular weight is about 25,000. D) Phenylalanine can be specifically attached to an —OH group at the 3' end. E) The tRNA must contain the sequence UUU.

E) The tRNA must contain the sequence UUU.

Which of the following is not true of tRNA molecules? A) The 3'-terminal sequence is —CCA. B) Their anticodons are complementary to the triplet codon in the mRNA. C) They contain more than four different bases. D) They contain several short regions of double helix. E) With the right enzyme, any given tRNA molecule will accept any of the 20 amino acids.

E) With the right enzyme, any given tRNA molecule will accept any of the 20 amino acids.

Posttranslational glycosylation of proteins is inhibited specifically by: A) chloramphenicol. B) cycloheximide. C) puromycin. D) streptomycin. E) tunicamycin.

E) tunicamycin.

The recognition of an amino acid by its cognate aminoacyl-tRNA synthetase is said to involve a "second genetic code". What is meant by this?

Each of the 20 amino acids has a unique synthetase, but many have multiple cognate tRNAs that must be charged (aminoacylated). The "second code" refers to features common to all tRNAs that carry the same amino acid, making them specifically recognizable by the correct synthetase. These features occur at different places in different tRNA classes, and may be as simple as a single G-U basepair, or require ten or more specific nucleotides throughout the sequence.

In no more than three sentences, describe a nonsense suppressor tRNA and how it differs from a normal tRNA.

A suppressor tRNA has one or more altered bases in its anticodon, which allows it to base pair with one of the stop codons. This allows the insertion of an amino acid instead of the chain termination that would normally have occurred at the point of a nonsense (stop) codon.

Aminoacyl-tRNA synthetases (amino acid activating enzymes): A) "recognize" specific tRNA molecules and specific amino acids. B) in conjunction with another enzyme attach the amino acid to the tRNA. C) interact directly with free ribosomes. D) occur in multiple forms for each amino acid. E) require GTP to activate the amino acid.

A) "recognize" specific tRNA molecules and specific amino acids.

Which of the following statements about tRNA molecules is false? A) A, C, G, and U are the only bases present in the molecule. B) Although composed of a single strand of RNA, each molecule contains several short, double-helical regions. C) Any given tRNA will accept only one specific amino acid. D) The amino acid attachment is always to an A nucleotide at the 3' end of the molecule. E) There is at least one tRNA for each of the 20 amino acids.

A) A, C, G, and U are the only bases present in the molecule.

Formation of the ribosomal initiation complex for bacterial protein synthesis does not require: A) EF-Tu. B) formylmethionyl tRNAfMet. C) GTP. D) initiation factor 2 (IF-2). E) mRNA.

A) EF-Tu.

Glycosylation of proteins inside the endoplasmic reticulum does not involve: A) a His residue on the protein. B) an Asn residue on the protein. C) dolichol phosphate. D) glucose. E) N-acetylglucosamine.

A) a His residue on the protein.

Describe the sequence of events between the transcription of an mRNA for a secreted protein and the arrival of that protein in the lumen of the endoplasmic reticulum.

(1) The mRNA forms an initiation complex with a cytoplasmic ribosome and transcription begins. (2) The amino-terminal portion of the nascent chain, containing a signal sequence, binds to an SRP (signal recognition particle), interrupting polypeptide elongation. (3) The SRP-ribosome-nascent chain complex binds to the ribosome and SRP receptors on the cytosolic face of the ER; the SRP then dissociates. (4) Chain elongation continues, with the newly synthesized polypeptide crossing into the ER lumen as it grows. (5) The signal sequence is cleaved.

Describe, succinctly, two ways in which synthetic polynucleotides were used in solving the genetic code (you need not describe how the synthetic polynucleotides were made).

(1) When synthetic polymers of only one nucleotide were used as mRNA in vitro, only one of the 20 amino acids was converted into protein. For example, poly(U) (containing only the codon UUU) directed the synthesis of polyphenylalanine, showing that UUU encodes Phe. (2) Trinucleotides of known sequence were used to stimulate aminoacyl-tRNA binding to ribosomes. Because only that aminoacyl-tRNA whose anticodon matched the trinucleotide "mRNA" was bound, the coding specificity of each sequence of three bases could be determined by determining which of the 20 aminoacyl-tRNAs bound. (3) Random polymers of RNA containing known ratios of nucleotides (e.g., 70% A and 30% T) generate only certain codons in predictable ratios. The identities and ratios of the amino acids specified by such polymers provided important clues that helped solve the genetic code. (4) Additional assignments were made possible using synthetic oligonucleotides containing repeats of specific two, three, or four base pair sequences.

The following sequence of four amino acids occurred in the structure of a polypeptide found in a wild-type organism: Leu-Ser-Ile-Arg. Several mutants were isolated, each of which carried a single base pair change in the region of DNA that coded for this amino acid sequence. Their corresponding amino acid sequences are: Mutant 1 MET-Ser-Ile-Arg 2 Leu-TRP-Ile-Arg 3 Leu-Ser-ARG-Arg 4 Leu-Ser-Ile-PRO 5 Leu-Ser-Ile-TRP What was the nucleotide sequence of the region of mRNA that coded for the amino acid sequence in the wild-type organism?

(5')C or (5')U UG UCG AUA CGG

The template strand of a segment of double-stranded DNA contains the sequence: (5')CTT TGA TAA GGA TAG CCC TTC (a) What is the base sequence of the mRNA that can be transcribed from this strand? (b) What amino acid sequence could be coded by the mRNA base sequence in (a), using only the first reading frame starting at the 5' end? (Refer to Fig. 27-7, p. 1069.) (c) Suppose the other (complementary) strand is used as a template for transcription. What is the amino acid sequence of the resulting peptide, again starting from the 5' end and using only the first reading frame?

(a) (5')GAA GGG CUA UCC UUA UCA AAG(3') (b) Glu-Gly-Leu-Ser-Leu-Ser-Lys (c) The codons translate to Leu-Stop-Stop. No peptide would be produced because of the stop codons.

Briefly describe the role of the following components in bacterial protein synthesis. (a) Initiation factor 2 (IF-2) (b) 16S RNA (c) Peptidyl transferase (d) Release factors (e) Elongation factor G (EF-G) (f) N10-formyltetrahydrofolate (g) ATP (h) tRNAfMet

(a) IF-2 is a protein factor that, when bound to GTP, brings the fMet-tRNAfMet to the initiation complex. (b) 16S RNA is a component of the small (30S) subunit. It contains a sequence complementary to the Shine-Dalgarno sequence in the mRNA, and helps to line up the mRNA initiation AUG codon on the ribosome. (c) Peptidyl transferase is a ribozyme in the 50S ribosomal subunit. It catalyzes formation of each peptide bond as the ribosome moves along the mRNA. (d) Release factors are proteins that bring about the release of the finished polypeptide when the ribosome encounters a termination codon in the mRNA. (e) EF-G participates in the translocation of the ribosome down the mRNA by one codon after each peptide bond is formed. (f) N10-formyl-tetrahydrofolate is the cofactor that donates a methyl group in the conversion of tRNA-bound Met to fMet. (g) ATP is the substrate for aminoacyl-tRNA synthetases; it donates an AMP residue in the formation of aminoacyl adenylate, which donates the aminoacyl group to tRNA. (h) tRNAfMet is the transfer RNA that initiates protein synthesis by inserting the first amino acid (fMet) in every prokaryotic protein.

You have isolated a fragment of viral DNA that totally encodes at least two proteins, 120 and 80 amino acids long. The DNA fragment is 400 base pairs long. (a) Why might you consider this unusual? (b) You sequence the two proteins and find no sequence homology. Propose a model to account for these findings.

(a) Two distinct proteins of these sizes should require mRNAs of 360 and 240 base pairs because each amino acid residue requires 3 base pairs to code for it. (b) No homology means that the smaller protein cannot be derived from the larger by proteolysis; if it were, there would be 80 amino acid residues of identical sequence in the two proteins. One possible explanation is that the two genes coding for these proteins overlap and are read in different reading frames.

Following the synthesis of their polypeptide chain, many proteins require further posttranslational modifications before they attain their full biological activity or function. List and describe briefly at least four possible types of modification that can occur.

1) Amino-terminal modification (N-acetylation or deacetylation), 2) removal of signal sequences used for targeting, 3) side-chain modification (phosphorylation, carboxylation, methylation, etc.), 4) attachment of N-linked (to Asn) or O-linked (to Ser or Thr) oligosaccharide moieties, 5) isoprenylation of Cys side-chains, 6) incorporation of prosthetic groups (heme, biotin, etc.), 7) processing of proenzymes or zymogens, and 8) formation of disulfide crosslinks.

Number the following steps in the proper order with regard to protein synthesis. ___ Aminoacyl-tRNA binds to the A site. ___ Deacylated tRNA is released from ribosome. ___ Peptide bond formation shifts the growing peptide from the P to the A site. ___ The 50S subunit binds to the initiation complex of the 30S subunit and mRNA.

2; 4; 3; 1

Which of the following is(are) true for protein synthesis in eukaryotes? A) All proteins are initially synthesized with methionine at their C-terminus. B) All proteins are initially synthesized with methionine at their N-terminus. C) All proteins are initially synthesized with tryptophan at their C-terminus. D) All proteins are initially synthesized with a multiple of 3 amino acids in their sequence. E) None of the above.

B) All proteins are initially synthesized with methionine at their N-terminus.

In E. coli, aminoacyl-tRNA synthetases: A) activate amino acids in 12 steps. B) are amino acid-specific; there is at least one enzyme specific for each amino acid. C) fall into two classes, each of which attaches amino acids to different ends of the tRNA. D) have no proofreading activities. E) require a tRNA, an amino acid, and GTP as substrates.

B) are amino acid-specific; there is at least one enzyme specific for each amino acid.

Bacterial ribosomes: A) bind tightly to specific regions of DNA, forming polysomes. B) contain at least one catalytic RNA molecule (ribozyme). C) contain three species of RNA and five different proteins. D) have specific, different binding sites for each of the 20 tRNAs. E) require puromycin for normal function.

B) contain at least one catalytic RNA molecule (ribozyme).

The large structure consisting of a mRNA molecule being translated by multiple copies of the macromolecular complexes that carry out protein synthesis is called a: A) lysosome. B) polysome. C) proteosome. D) ribosome. E) synthosome.

B) polysome.

Assuming that the average amino acid residue contributes 110 to the peptide molecular weight, what will be the minimum length of the mRNA encoding a protein of molecular weight 50,000? A) 133 nucleotides B) 460 nucleotides C) 1,400 nucleotides D) 5,000 nucleotides E) A minimum length cannot be determined from the data given.

C) 1,400 nucleotides

The pathway for polypeptides exported from E. coli includes the following steps, which occur in what order for correct export? 1. A chaperone, SecA, binds to the polypeptide. 2. A chaperone, SecB, binds to the polypeptide. 3. ATP is hydrolyzed by Sec A. 4. SecA pushes 20 amino acids of the polypeptide into the translocation complex. A) 1, 2, 3, 4 B) 1, 2, 4, 3 C) 2, 1, 4, 3 D) 2, 3, 1, 4 E) 3, 1, 4, 2

C) 2, 1, 4, 3

A certain bacterial mRNA is known to represent only one gene and to contain about 800 nucleotides. If you assume that the average amino acid residue contributes 110 to the peptide molecular weight, the largest polypeptide that this mRNA could code for would have a molecular weight of about: A) 800. B) 5,000. C) 30,000. D) 80,000. E) An upper limit cannot be determined from the data given.

C) 30,000.

Which of the following statements about bacterial mRNA is true? A) A ribosome usually initiates translation near the end of the mRNA that is synthesized last. B) An mRNA is never degraded but is passed on to the daughter cells at cell division. C) During polypeptide synthesis, ribosomes move along the mRNA in the direction 5' to 3'. D) Ribosomes cannot initiate internally in a polycistronic transcript. E) The codon signaling peptide termination is located in the mRNA near its 5' end.

C) During polypeptide synthesis, ribosomes move along the mRNA in the direction 5' to 3'.

Which one of the following antibiotics does not function by interfering with the translational process? A) Chloramphenicol B) Cycloheximide C) Penicillin D) Puromycin E) Streptomycin

C) Penicillin

Which one of the following is true about the genetic code? A) All codons recognized by a given tRNA encode different amino acids. B) It is absolutely identical in all living things. C) Several different codons may encode the same amino acid. D) The base in the middle position of the tRNA anticodon sometimes permits "wobble" base pairing with 2 or 3 different codons. E) The first position of the tRNA anticodon is always adenosine.

C) Several different codons may encode the same amino acid.

It is possible to convert the Cys that is a part of Cys-tRNACys to Ala by a catalytic reduction. If the resulting Ala-tRNACys were added to a mixture of (1) ribosomes, (2) all the other tRNAs and amino acids, (3) all of the cofactors and enzymes needed to make protein in vitro, and (4) mRNA for hemoglobin, where in the newly synthesized hemoglobin would the Ala from Ala-tRNACys be incorporated? A) Nowhere; this is the equivalent of a nonsense mutation B) Wherever Ala normally occurs C) Wherever Cys normally occurs D) Wherever either Ala or Cys normally occurs E) Wherever the dipeptide Ala-Cys normally occurs

C) Wherever Cys normally occurs

In protein synthesis, 61 codons specify the 20 amino acids. Base pairing between the codon and the tRNA anticodon assures that the correct amino acid will be inserted into the nascent polypeptide chain. Why then does the cell require only 32 different tRNAs to recognize 61 different codons?

Certain tRNAs have the unusual nucleotide inosinate in the first anticodon position. Because inosinate can base pair with A, U, or C, a tRNA containing hypoxanthine can recognize three different codons. In each recognized codon, there is a standard anticodon-codon base pair with the first two bases of the codon; "wobble" in the third base pair allows one tRNA to read three different codons. Similarly, tRNAs with U or G in the first anticodon position also exhibit a wobble effect that permits pairing with two different codons.

Approximately how many NTPs must be converted to NDPs to incorporate one amino acid into a protein? A) 0 B) 1 C) 2 D) 4 E) 8

D) 4

In bacteria the elongation stage of protein synthesis does not involve: A) aminoacyl-tRNAs. B) EF-Tu. C) GTP. D) IF-2. E) peptidyl transferase.

D) IF-2.

Which of the following are features of the wobble hypothesis? A) A naturally occurring tRNA exists in yeast that can read both arginine and lysine codons. B) A tRNA can recognize only one codon. C) Some tRNAs can recognize codons that specify two different amino acids, if both are nonpolar. D) The "wobble" occurs only in the first base of the anticodon. E) The third base in a codon always forms a normal Watson-Crick base pair.

D) The "wobble" occurs only in the first base of the anticodon.

Which of the following statements about aminoacyl-tRNA synthetases is false? A) Some of the enzymes have an editing/proofreading capability. B) The enzyme attaches an amino acid to the 3' end of a tRNA. C) The enzyme splits ATP to AMP + PPi. D) The enzyme will use any tRNA species, but is highly specific for a given amino acid. E) There is a different synthetase for every amino acid.

D) The enzyme will use any tRNA species, but is highly specific for a given amino acid.

Which of the following is true about the sorting pathway for proteins destined for incorporation into lysosomes or the plasma membrane of eukaryotic cells? A) Binding of SRP to the signal peptide and the ribosome temporarily accelerates protein synthesis. B) The newly synthesized polypeptides include a signal peptide at their carboxyl termini. C) The signal peptide is cleaved off inside the mitochondria by signal peptidase. D) The signal recognition particle (SRP) binds to the signal peptide soon after it appears outside the ribosome. E) The signal sequence is added to the polypeptide in a posttranslational modification reaction.

D) The signal recognition particle (SRP) binds to the signal peptide soon after it appears outside the ribosome.

Which one of the following statements about ribosomes is true? A) The large subunit contains rRNA molecules, the small subunit does not. B) The RNA in ribosomes plays a structural, not catalytic, role. C) There are about 25 of them in an E. coli cell. D) There are two major subunits, each with multiple proteins. E) They are relatively small, with molecular weights less than 10,000.

D) There are two major subunits, each with multiple proteins.

The enzyme that attaches an amino acid to a tRNA (aminoacyl-tRNA synthetase): A) always recognizes only one specific tRNA. B) attaches a specific amino acid to any available tRNA species. C) attaches the amino acid at the 5' end of the tRNA. D) catalyzes formation of an ester bond. E) splits ATP to ADP + Pi.

D) catalyzes formation of an ester bond.

The signal sequences that direct proteins to the nucleus are: A) always at the amino terminus of the targeted protein. B) cleaved after the protein arrives in the nucleus. C) glycosyl moieties containing mannose 6-phosphate residues. D) not located at the ends of the peptide, but in its interior. E) the same as those that direct certain proteins to lysosomes.

D) not located at the ends of the peptide, but in its interior.

Ubiquitin-mediated protein degradation is a complex process, and many of the signals remain unknown. One known signal involves recognition of amino acids in a processed protein that are either stabilizing (Ala, Gly, Met, Ser, etc.) or destabilizing (Arg, Asp, Leu, Lys, Phe, etc.), and are located at: A) a helix-turn-helix motif in the protein. B) a lysine-containing target sequence in the protein. C) a zinc finger structure in the protein. D) the amino-terminus of the protein. E) the carboxy-terminus of the protein.

D) the amino-terminus of the protein.

In the "activation" of an amino acid for protein synthesis: A) leucine can be attached to tRNAPhe, by the aminoacyl-tRNA synthetase specific for leucine. B) methionine is first formylated, then attached to a specific tRNA. C) the amino acid is attached to the 5' end of the tRNA through a phosphodiester bond. D) there is at least one specific activating enzyme and one specific tRNA for each amino acid. E) two separate enzymes are required, one to form the aminoacyl adenylate, the other to attach the amino acid to the tRNA.

D) there is at least one specific activating enzyme and one specific tRNA for each amino acid.

Indicate whether each of the following statements is true (T) or false (F). ___Assembly of a complete ribosome onto an mRNA requires ATP hydrolysis. ___Aminoacylation or "charging" of tRNA requires the formation of an aminoacyl-AMP intermediate. ___Aminoacyl-tRNA binding to the A site of the ribosome requires the accessory factor EF-G and GTP hydrolysis. ___Translocation of a growing polypeptide from the A to the P site on the ribosome requires EF-G and GTP hydrolysis. ___Termination of translation requires release factors, but no NTP hydrolysis.

F; T; F; T; T

A given mRNA sequence might be translated in any of three reading frames. Describe how prokaryotes and eukaryotes determine the correct reading frame.

In prokaryotes, the Shine-Dalgarno sequence in the mRNA base pairs with a complementary sequence in the 16S RNA of the ribosome; this positions the correct start codon (AUG) on the 30S ribosomal subunit. Thus, the initiating AUG is distinguished by its proximity to the Shine-Dalgarno sequence. In eukaryotes, the initiating AUG codon is the first AUG that the ribosome encounters as it scans the mRNA from its 5' end. In both cases the initiating AUG also sets the correct reading frame.

What are the stages in targeting of nuclear proteins, and why are the targeting sequences not removed upon arrival of the protein in the nucleus?

In the cytoplasm, where eukaryotic protein synthesis occurs, proteins carrying nuclear localization signal (NLS) sequences are bound by a complex of importin and , which is then bound to a nuclear pore. Ran GTPase mediates translocation of this complex into the nucleus, where importin dissociates from importin , and importin releases the nuclear protein. Importin and are then exported from the nucleus, and available for another cycle of import. The nuclear envelope of higher eukaryotes breaks down at each cell division, distributing the nuclear contents throughout the cell. When the nuclear envelope is reestablished, NLS-carrying proteins can be reimported to fulfill their function.

In 1961, Howard Dintzis carried out an experiment that defined the direction of polypeptide chain growth during protein synthesis in cells. The experiment involved the analysis of hemoglobin molecules that were being synthesized in reticulocytes in the presence of radioactive amino acids. Describe the analysis and how it demonstrated the direction of chain growth.

In the experiment, only completed polypeptides were isolated and analyzed for the amount of redioactivity in different regions. The greatest radioactivity should be located in the region furthest from the initiation point and the least radioactivity in the region closest to the initiation point. The opposite is true for the termination point. The analysis showed that polypeptide synthesis was initiated at the amino terminus and terminated at the carboxyl terminus.

Consider the following hypothetical short mRNA; what would be the sequence of the protein produced if this were translated in an E. coli cell? 5'-AUAGGAGGUUUGACCUAUGCCUCGUUUAUAGCC-3'

N-met-pro-arg-leu-C

Regarding translation in eukaryotes versus that in prokaryotes (bacteria), indicate whether each of the following statements is true (T) or false (F). ___ In eukaryotes the 3' end of the mRNA is associated with the 5' end during initiation whereas in prokaryotes it is not. ___ In prokaryotes it is initiated at an AUG near a Shine-Dalgarno sequence in the mRNA whereas in eukaryotes it is initiated at an AUG near the 3' end of the mRNA. ___ In prokaryotes it is initiated with Met whereas in eukaryotes it is initiated with fMet. ___ In prokaryotes translation and transcription are coupled whereas in eukaryotes they are not.

T, F, F, T

Indicate whether each of the following statements is true (T) or false (F). ___ Bacterial mRNA is broken down within a few minutes of its formation in E. coli. ___ Bacterial mRNA consists only of the bases that code for amino acids. ___ Polysomes do not necessarily contain mRNA. ___ Bacterial mRNA normally occurs as a double-stranded structure, with one strand containing codons, the other containing anticodons. ___ Bacterial mRNA can be translated while it is still being synthesized.

T; F; F; F; T

Indicate whether the following statements are true (T) or false (F). ___A ribosome is the complex within which protein synthesis occurs. ___Ribosomes contain many separate proteins. ___The three ribosomal RNAs in a bacterial ribosome are distributed in three separate, large ribosomal subunits. ___There are four binding sites for aminoacyl-tRNAs on a ribosome.

T; T; F; F

A new antibiotic was recently discovered that inhibits prokaryotic protein synthesis. In the presence of the antibiotic, protein synthesis can be initiated, but only dipeptides that remain bound to the ribosome are formed. What specific step of protein synthesis is likely to be blocked by this antibiotic?

The antibiotic probably blocks translocation.

When first synthesized, proinsulin has an additional leader or signal peptide at its amino terminus. This complete molecule is called preproinsulin and the signal peptide is cleaved off to give proinsulin. Briefly, what is the likely function of the signal peptide?

The leader peptide in proinsulin is a signal sequence directing it to the endoplasmic reticulum, from which it enters the Golgi complex and is packaged into a secretory vesicle for secretion by exocytosis.

Describe the possible outcomes that could occur because of a single base change in an mRNA

The most likely result is a single amino acid change in the encoded protein, when a codon is altered to one of another amino acid. However, some nucleotide changes will be "silent" and not change the protein, if the altered codon still specifies the original amino acid. Conversion to a nonsense codon will result in a truncated polypeptide, while alteration of the normal stop codon to a "sense" codon will result in a lengthened protein. Alternation of the initiaton codon may result in the total failure to translate, and result in no protein product. Changes in mRNA sequence outside the protein-coding region may affect translational efficiency, splicing, or mRNA turnover rates.

Polypeptide chain elongation in E. coli occurs by the cyclical repetition of three steps. What are these steps and what cellular components are necessary for each of them to occur?

The three steps are: (1) An aminoacyl-tRNA is brought to the A site by EF-Tu with bound GTP; (2) peptidyl transferase (a ribozyme) catalyzes peptide-bond formation; (3) the ribosome translocates three nucleotides down the mRNA, helped by EF-G (translocase). This shifts the peptidyl-tRNA to the A site and the deacylated tRNA to the E site.

The process of charging tRNAs with their cognate amino acids involves multiple proofreading steps to increase the overall fidelity. Briefly describe these steps.

There are two main stages of selection: 1) the synthetase strongly favors activation of the correct amino acid to become aminoacyl-AMP (incorrect amino acids are very poorly activated) and 2) when the correct uncharged tRNA is bound to the enzyme, only the correct aminoacyl-AMP is tolerated in the "proofreading" active site (incorrect aminoacyl-AMPs, though they may become bound, are rapidly hydrolyzed). In addition, for most synthetases, if their tRNA does manage to become acylated by the wrong amino acid, that product is also rapidly hydrolyzed.

Describe the role of ubiquitin in mediating intracellular protein breakdown.

Ubiquitin, a protein found in all eukaryotic cells, is covalently joined to proteins targeted for degradation. The carboxyl-terminal residue of ubiquitin is first activated by the formation of a thioester with the first enzyme in the pathway in an ATP-dependent reaction. After displacement of the first enzyme by a second, also yielding a thioester link, ubiquitin is finally joined through its carboxyl terminus to a lysine -amino group in the protein to be degraded. The presence of ubiquitin targets the protein for degradation by cellular proteases.

Outline one of the experimental methods providing evidence that the genetic code was a triplet code.

When one or two nucleotides were added to or deleted from a gene, the resulting mRNA produced a protein with a different amino acid sequence after the deletion or insertion. When three nucleotides were added or deleted, the resulting protein had a normal sequence except for the insertion or deletion of a single amino acid residue.