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May 9, 2016
Week 7
Study Questions from the textbook:
Chapter 12- 12.5, 12.6, 12.10, 12.13, 12.20, 12.25, 12.34
12.5 (a) Why is the genetic code a triplet code instead of a singlet or doublet code?
(b) How many different amino acids are specified by the genetic code?
(c) How many different amino acid sequences are possible in a polypeptide 146 amino
acids long?
ANS: (a) Singlet and doublet codes provide a maximum of 4 and (4)2 or 16 codons,
respectively. Thus neither code would be able to specify all 20 amino acids. (b) 20. (c)
(20)146.
12.6
What types of experimental evidence were used to decipher the genetic code?
ANS: Synthetic RNA molecules (polyuridylic acid molecules) containing only the base
uracil were prepared. When these synthetic molecules were used to activate in vitro
protein synthesis systems, small polypeptide containing only the amino acid
phenylalanine (polyphenylalanine molecules) were synthesized. Codons composed only
of uracil were thus shown to specify phenylalanine. Similar experiments were carried out
using synthetic RNA molecules with different base compositions. Later, in vitro systems
activated with synthetic RNA molecules with known repeating base sequences were
developed. Ultimately, in vitro systems in which specific aminoacyl-tRNAs where shown
to bind to ribosomes activated with specific mini-mRNAs, which were trinucleotides of
known base sequence, were developed and used in codon identification.
12.10. What is the minimum number of tRNAs required to recognize the six codons
specifying the amino acid leucine?
ANS: Because of wobble (Table 12.2), one tRNA can recognize both UUA and UUG
codons for leucine. However, it takes two more tRNAs to recognize CUU, CUC, CUA,
and CUG. Therefore, a minimum of three tRNAs are required to recognize the six
codons for leucine.
12.13 Identify three different types of RNA that are involved in translation and list the
characteristics and functions of each.
ANS: Messenger RNA (mRNA) molecules carry genetic information from the
chromosomes (where the information is stored) to the ribosomes in the cytoplasm (where
the information is expressed during protein synthesis). The linear sequence of triplet
codons in an mRNA molecule specifies the linear sequence of amino acids in the
polypeptide(s) produced during translation of that mRNA. Transfer RNA (tRNA)
molecules are small (about 80 nucleotides long) molecules that carry amino acids to the
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b) Currently, the E site is empty, the P site contains an uncharged tRNA, and the A site
contains the growing polypeptide-tRNA. Next, the peptidyl tRNA in the A site of the
ribosome will be translocated to the P site, and the uncharged tRNA in the P site will be
translocated to the E site, as the ribosome moves 3 nucleotides toward the 3 end of the
mRNA. The translocation of the ribosome is catalyzed by EF-G and the energy of GTP
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hydrolysis. The translocation of the peptidyl tRNA from the A site to the P site leaves the
A site unoccupied. As the next codon is GCC, the A site will next bind tRNAAla.
*2. An experiment by Khorana and his colleagues translated a synthetic mRNA
containing repeats of the trinucleotide UUG (UUGUUGUUGUUGUUG).
a) How many (triplet, nonoverlapping) reading frames are possible in this mRNA?
b) What is the result obtained from each reading frame?
c) How does the result of this experiment help confirm the triplet nature of the genetic
code?
ANS: a) Three reading frames are possible (5-UUG UUG UUG-3, 5-UGU UGU
UGU-3, and 5-GUU GUU GUU-3).
b) translation of 5-UUG UUG UUG-3 produces polyleucine (Leu). Translation of 5UGU UGU UGU-3 produces polycystine (Cys). Translation of 5-GUU GUU
GUU-3 produces polyvaline (Val).
c) The result differs from that predicted for nonoverlapping doublet or quadruplet codes,
both of which predict a single type of polypeptide composed of repeats of two, or three
amino acids. Only a triplet code is predicted to produce three different single amino acid
polypeptides from the given mRNA.
Chapter 13- 13.2, *13.15, 13.16, 13.22, *13.26, 13.28, 13.29, 13.31, 13.37
13.2. Of all possible missense mutations that can occur in a segment of DNA encoding
the amino acid tryptophan, what is the ratio of transversions to transitions if all single
base-pair substitutions occur at the same frequency?
ANS: 6:1
UGG transitions UGA (nonsense), UAG (nonsense), CGG (Arg).
UGG transversions UGC (Cys), UGU (Cys), UCG (Ser), UUG (Leu), AGG (Arg), GGG
(Gly).
13.15 If CTT is a DNA triplet (transcribed strand of DNA) specifying glutamic acid,
what DNA and mRNA base triplet alterations could account for valine and lysine in
position 6 of the -globin chain?
ANS:
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13.16. The bacteriophage T4 genome contains about 50 percent A:T base pairs and 50
percent G:C base pairs. The base analog 2-aminopurine induces A:T G:C and G:C
A:T base-pair substitutions by undergoing tautomeric shifts. Hydroxylamine is a
mutagenic chemical that reacts specifically with cytosine and induces only G:C A:T
substitutions. If a large number of independent mutations were produced in bacteriophage
T4 by treatment with 2-aminopurine, what percentage of these mutations should you
expect to be induced to mutate back to the wild-type genotype by treatment with
hydroxylamine?
ANS: About half of the induced mutations would be expected to mutate back to the wildtype genotype.
13.22. One person was in an accident and received 50 roentgens (r) of X rays at one
time. Another person received 5 r in each of 20 treatments. Assuming no intensity effect,
what proportionate number of mutations would be expected in each person?
ANS: The person receiving a total of 100 r would be expected to have twice as many
mutations as the one receiving 50r.
*13.26. You are screening three new pesticides for potential mutagenicity using the Ames
test. Two his strains resulting from either a frameshift or a transition mutation were used
and produced the following results (number of revertant colonies):
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13.31. Mutations in the genes encoding the and subunits of hemoglobin lead to
blood diseases such as thalassemias and sickle-cell anemia. You have found a family in
China in which some members suffer from a new genetic form of anemia. The DNA
sequences at the 5 end of the nontemplate strand of the normal and mutant DNA
encoding the subunit of hemoglobin are as follows:
Normal 5-ACGTTATGCCGTACTGCCAGCTAACTGCTAAAGAACAATTA-3
Mutant 5-ACGTTATGCCCGTACTGCCAGCTAACTGCTAAAGAACAATTA.-3
(a) What type of mutation is present in the mutant hemoglobin gene?
(b) What are the codons in the translated portion of the mRNA transcribed from the
normal and mutant genes?
(c) What are the amino acid sequences of the normal and mutant polypeptides?
ANS: (a) Frameshift due to the insertion of C at the 9th, 10th, or 11th nucleotide from
the 5 end.
(b) Normal: 5-AUGCCGUACUGCCAGCUAACUGCUAAAGAACAAUUA-3.
Mutant: 5-AUGCCCGUACUGCCAGCUAACUGCUAAAGAACAAUUA-3.
(c) Normal: NH2-Met-Pro-Tyr-Cys-Gln-Leu-Thr-Ala-Lys-Glu-Gln-Leu.
Mutant: NH2-Met-Pro-Val-Leu-Pro-Ala-Asn-Cys.
13.37. Would you expect nitrous acid to induce a higher frequency of Tyr Ser or Tyr
Cys substitutions? Why?
ANS: Tyr Cys substitutions; Tyr to Cys requires a transition, which is induced by
nitrous acid. Tyr to Ser would require a transversion, and nitrous acid is not expected to
induce transversions.
Study Questions from the literature:
Crick et al, 1961.
1. Outline the experiment and major results of this study in your own words.
ANS: As stated in the question, this should be in your own words.
They began with a T4 phage mutant for the rII gene, where the mutation caused a single
base-pair addition, and thus this phage was unable to infect bacteria and make plaques.
They added additional mutations to this original mutant phage, which caused single base
pair insertions or deletions, and examined whether and in what combination additional
these additional mutations were able cause a reversion (i.e. restore wild type function) in
the phage. What they observed was that adding a single deletion to their original mutant
with a single insertion caused reversion. Adding an additional insertion to their original
mutant with an insertion (for a total of 2bp inserted) did not cause reversion. When they
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added two additional insertions to their original insertion (for a total of 3bp inserted) they
observed a reversion.
From these experiments, they were able to observe that:
- The genetic code is a triplet code. Three base pairs in the mRNA specify an amino
acid. Because an addition of a total of 3 base pairs is able to restore wild type function of
the phage (where as the addition of one or two bases is not) the code must work in threes.
-The bases are read from a fixed starting point, and there is no punctuation to
indicate what the right codons are. Because the addition of one base the reading into
triplets is incorrect, meaning that the reading frame is thrown off, thus causing a
frameshift.
From previous studies, they knew:
- The genetic code is not overlapping. A mutation in a gene that affects only one base
pair in the DNA only affects one amino acid in the protein encoded by that gene
From simple calculations they propose:
- The genetic code is degenerate. They make the observation that there are 64
combinations of the four DNA bases in a three base pair codon (4 x 4 x 4). There are only
20 amino acids. Therefore, it must be that each amino acid is specified by more than one
codon.
2. If the genetic code had turned out to be a non-overlapping quadruplet code (4 bases per
amino acid), how would their results have changed?
ANS: Their experiments started with a T4 phage with a single base pair insertion in the
rII gene, which had an rII- phenotype, and added additional insertions or deletions to that
strain. If the genetic code was a quadruplet code, they would not have observed reversion
to rII+ with the two additional insertions (for a total of 3bp inserted). Instead, it would
have taken 3 additional insertions (for a total of 4 bp inserted) to restore the reading
frame and the rII+ phenotype.