Sample Examination Questions for Exam 3 Material - Solved Biology 3300 / Dr. Jerald Hendrix Warning! These questions are posted solely to provide examples of past test questions. There is no guarantee that any of these questions will be on any examination in the future. Students are responsible for all of the material covered in lectures, assigned readings, textbook problems, laboratories, and any other assigned work. Since these samples have been taken from several past exams, some questions may be very similar or identical. On short answer, essay questions, and genetics problems, the point values from previous exams have been included to give an indication of approximately how much “weight” was given to a question in the past; however, there is no guarantee that any particular question, format, or point distribution will be used on any examination. 5. Which of the following enzymes catalyze the attachment of an amino acid to tRNA in the formation of aminoacyl tRNA? (a) (b) (c) (d) (e) 15. In the formation of an aminoacyl tRNA (a) (b) (c) (d) (e) 18. RNA polymerase aminoacyl tRNA synthetase Type I restriction endonuclease Type II restriction endonuclease reverse transcriptase the amino terminus of the amino acid is directly attached to the 5' end of the tRNA. the carboxyl terminus of the amino acid is directly attached to the 5' end of the tRNA. the amino terminus of the amino acid is directly attached to the 3' end of the tRNA. the carboxyl terminus of the amino acid is directly attached to the 3' end of the tRNA. the side chain group of the amino acid is directly attached to the anticodon loop of the tRNA. Which of the following statements is true? (a) (b) (c) (d) (e) Ribosomes read mRNA from the 5' to the 3' end and synthesize the nascent protein chain from the carboxyl to the amino terminus. Ribosomes read mRNA from the 3' to the 5' end and synthesize the nascent protein chain from the amino to the carboxyl terminus. Ribosomes read mRNA from the 5' to the 3' end and synthesize the nascent protein chain from the amino to the carboxyl terminus. Ribosomes read mRNA from the 3' to the 5' end and synthesize the nascent protein chain from the carboxyl to the amino terminus. Ribosomes read DNA from the 5' to the 3' end and synthesize the nascent RNA chain from the 3' to the 5' end. Page 1 22. In prokaryotic genes, a promoter consensus sequence located about 10 bases upstream from the start of transcription is the (a) (b) (c) (d) (e) 24. CAAT box. Hogness box. Shine-Dalgarno box. Meselson-Stahl box. Pribnow box. A promoter (a) (b) (c) (d) (e) is a protein that binds to RNA polymerase to enhance the process of transcription. is a DNA sequence to which RNA polymerase binds. is a DNA sequence to which Type I or Type II restriction endonucleases bind. is the location where DNA replication begins. is an initiation factor in the process of translation. 26. Energy for the process of elongation during translation comes from the hydrolysis of (a) ATP. (b) CTP. (c) GTP. (d) TTP. (e) UTP. 30. The enzyme responsible for forming the final phosphodiester bond between two DNA fragments during DNA replication is (a) (b) (c) (d) (e) 33. DNA ligase DNA polymerase I DNA polymerase III DNA-directed RNA polymerase reverse transcriptase Which of the following statements best describes the relationship between the nucleotide sequences of the template strand and the nontemplate strand in the DNA of a gene? (a) (b) (c) (d) (e) They are identical to each other. They are parallel to each other. They are covalently attached to each other. They are complementary to each other. U in the coding strand is found in place of the T in the noncoding stand. Page 2 38. Which of the following best describes the function of the stop codons UAA, UAG, and UGA? (a) (b) (c) (d) (e) 39 Okasaki fragments (a) (b) (c) (d) (e) 40. addition of a 5' methylated guanosine cap addition of a 3' poly-A tail splicing of RNA segments all of the above none of the above Amino acids are assembled into proteins (a) (b) (c) (d) (e) 1. are fragments of the discontinuous strand of nascent DNA during replication. are fragments of the continuous strand of nascent DNA during replication. are fragments of the template DNA during replication. are fragments of E. coli DNA polymerase I that have 5' exonuclease activity. are fragments of E. coli DNA polymerase I that have 3' exonuclease activity. Which of the following processes occurs to eukaryotic mRNA during posttranscriptional processing? (a) (b) (c) (d) (e) 48. Termination of DNA replication Origin of DNA replication Termination of transcription Termination of protein synthesis Orientation of the noncoding DNA strand of a gene inside the nucleus. on ribosomes. inside the Golgi apparatus. on the chromosomes. only during mitosis. In DNA replication, new nucleotides are added (a) (b) (c) (d) (e) to the 5' end of the each nascent strand. to the 3' end of each nascent strand. to both ends of each nascent strand. to the 5' end of the continuous strand and the 3' end of discontinuous strand fragments. to the 3' end of the continuous strand and the 5' end of discontinuous strand fragments. Page 3 2. RNA primase is responsible for (a) (b) (c) (d) (e) 3 The enzyme responsible for the most of the nascent chain synthesis during DNA replication in E. coli is (a) (b) (c) (d) (e) 4 the addition of new nucleotides to the 5' end of a nascent RNA strand during transcription. the addition of new nucleotides to the 3' end of a nascent RNA strand during transcription. the addition of the 5' leader sequence to eukaryotic mRNA during posttranscriptional processing. the addition of the 3' poly-A tail to eukaryotic mRNA during posttranscriptional processing. none of the above. DNA polymerase I DNA polymerase II DNA polymerase III DNA-directed RNA polymerase DNA ligase In an experiment, cells of actively growing E. coli were incubated for several minutes with radiolabeled deoxyribonucleosides (dNTPs). The DNA was isolated from the cells, denatured into single strands, and separated according to size by gradient centrifugation. The radioactivity in the tube corresponded to a single longer piece of DNA and a heterogeneous mixture of much shorter DNA fragments. Which of the following statements best explains these results? (a) (b) (c) (d) (e) The longer strand required a template, but the shorter strands did not. The longer strand had been synthesized from 5'→3', and the shorter fragments had been synthesized from 3'→5'. The longer strand had been synthesized from 3'→5', and the shorter fragments had been synthesized from 5'→3'. The longer strand represented discontinuous strand synthesis, and the shorter fragments represented continuous strand synthesis. The longer strand represented continuous strand synthesis, and the shorter fragments represented discontinuous strand synthesis. Page 4 5 Okasazi fragments (a) (b) (c) (d) (e) 6 The 3' exonuclease activity of DNA polymerase I in E. coli is responsible for (a) (b) (c) (d) (e) 41. are fragments of nascent DNA formed in discontinuous strand synthesis during DNA replication. are fragments of nascent DNA formed in continuous strand synthesis during DNA replication. are double-stranded DNA molecules with sticky ends, formed by cleavage with type II restriction endonucleases. are fragments of RNA that are spliced out and discarded during posttranslational processing of eukaryotic mRNA. are fragments of the amino-terminal end of a nascent protein chain, removed by post-translational proteolytic cleavage in almost all proteins. removal of the primer. removal of nucleotides with mismatched bases. removal of introns. addition of new nucleotides. formation of the final phosphodiester bond between two adjacent nascent chain fragments. Eukaryotic cells can be grown in a medium containing bromodeoxyuridine (BrdU), which substitutes for thymidine during DNA replication. A fluorescent dye that binds to double-stranded DNA makes it possible to recognize chromosomes or chromatids in which one or both strands of the DNA helix contain BrdU. The chromosome or chromatid fluoresces brightly when only one strand of its DNA contains BrdU, but it fluoresces weakly when both strands of DNA contain BrdU. In an experiment, unlabeled human cells in G1 were transferred to a medium containing BrdU and colchicine. The cells proceeded through one round of DNA replication, but the colchicine halted mitosis at metaphase. The metaphase chromosomes were isolated from the cells and analyzed under a fluorescence microscope. Assuming that DNA replication in humans is semiconservative, which of the following do you predict will best describe the appearance of the metaphase chromosomes? (a) (b) (c) (d) (e) One chromosome in each homologous pair will be brightly fluorescent, and the other chromosome will not be fluorescent. In every chromosome, each chromatid will be brightly fluorescent. In every chromosome, one chromatid will be weakly fluorescent, and the other chromatid will not be fluorescent. In every chromosome, each chromatid will be weakly fluorescent. In every chromosome, one chromatid will be weakly fluorescent and one chromatid will be brightly fluorescent. Page 5 7 A “shuttle vector” is a recombinant DNA vector capable of replicating in more than one bacterial species. Which of the following must a shuttle vector have? (a) (b) (c) (d) (e) Pribnow boxes from both bacterial species. Promoter regions from both bacterial species. Shine-Dalgarno sequences from both bacterial species. Origins of replication from both bacterial species. Ribosomes from both bacterial species. *********************************************************************** Eukaryotic cells can be grown in a medium containing bromodeoxyuridine (BrdU), which substitutes for thymidine during DNA replication. A fluorescent dye that binds to double-stranded DNA makes it possible to recognize chromosomes or chromatids with BrdU incorporated in their DNA. You can tell the difference between unlabeled chromatids, chromatids with one strand of the DNA double helix labeled, and chromatids with both strands of the DNA double helix labeled. The chromatid fluoresces brightly when only one strand contains BrdU, but weakly when both strands contain BrdU. The following diagrams represent the possible fluorescence patterns in metaphase chromosomes. Double hatch marks represent bright fluorescence, single marks represent weak fluorescence, and a clear white chromatid represents no fluorescence. Key: I. II. III. Neither chromatid is labeled with BrdU. (No fluorescence) Each chromatid is labeled in one strand of DNA. (Brightly fluorescent) Each chromatid is labeled in both strands of DNA. (Weakly fluorescent) Page 6 IV. V. VI. One chromatid is labeled in one strand only, and the other chromatid is labeled in both strands. One chromatid is unlabeled, and the other chromatid is labeled in one strand only. One chromatid is unlabeled, and the other chromatid is labeled in both strands. A group of cultured eukaryotic cells, all at the G1 stage and containing unlabeled chromosomes, were placed in a medium containing BrdU and allowed to grow. Samples were removed during the first, second, and third mitotic divisions. Mitosis in each of these samples was arrested at metaphase using colchicine. Chromosomes were isolated from cells in the samples and examined for fluorescence. Remember that chromosomes isolated at metaphase are in the replicated state (a pair of chromatids connected at the centromere). In addition, each chromatid in a replicated chromosome has one double helix. The two helices are the products of DNA replication. See next page for an explanation. 8 IF eukaryotic DNA replication were conservative, then what will be the appearance of the metaphase chromosomes after the third mitotic division? (The Roman numerals refer to the diagram above.) (a) (b) (c) (d) (e) 9 IF eukaryotic DNA replication were semiconservative, then what will be the appearance of the metaphase chromosomes after the third mitotic division? (The Roman numerals refer to the diagram above.) (a) (b) (c) (d) (e) 10 1/3 will look like I and 2/3 will look like III. 1/4 will look like II, 1/4 like III, 1/4 like V, and 1/4 like VI. 1/2 will look like III and 1/2 will look like IV. 3/4 will look like II and 1/4 will look like V. 3/4 will look like III and 1/4 will look like VI. 1/3 will look like I and 2/3 will look like III. 1/4 will look like II, 1/4 like III, 1/4 like V, and 1/4 like VI. 1/2 will look like III and 1/2 will look like IV. 3/4 will look like II and 1/4 will look like V. 3/4 will look like III and 1/4 will look like VI. Based on what you learned about DNA replication in class, what do you predict about the appearance of the metaphase chromosomes after the third mitotic division? (The Roman numerals refer to the diagram above.) (a) (b) (c) (d) (e) 1/3 will look like I and 2/3 will look like III. 1/4 will look like II, 1/4 like III, 1/4 like V, and 1/4 like VI. 1/2 will look like III and 1/2 will look like IV. 3/4 will look like II and 1/4 will look like V. 3/4 will look like III and 1/4 will look like VI. Page 7 The BrdU problem is similar to the Meselson-Stahl experiment. At the start of the experiment, each chromosome (DNA double helix) is unreplicated, and its strands are unlabeled.. The G1 cells are transferred to BrdU before the first round of replication begins, and the cells remain in BrdU for the remainder of the experiment. This means that every new strand formed will be labeled in BrdU. When the chromosomes are analyzed after the 1st, 2nd, & 3rd round of replication, each pair of sister chromatids represents the two daughter helices formed during replication in the S phase. Let a blue line | represent an unlabeled strand, and a red line | represent a labeled strand. If replication is CONSERVATIVE, the DNA during the experiment will follow this pattern: || Unlabeled, unreplicated helix at the start of the experiment ↓ st 1 round of replication. The two old strands stay together, and the two new strands join together if replication is conservative ||•|| All ↓ 2nd round of replication. ||•|| ||•|| rd ½ and ½ ↓ 3 round of replication. ||•|| ||•|| ||•|| ||•|| ¼ and ¾ , or ¼ like VI and ¾ like III. Page 8 If replication is SEMICONSERVATIVE, the DNA during the experiment will follow this pattern: || Unlabeled, unreplicated helix at the start of the experiment st ↓ 1 round of replication. In semiconservative replication, each new helix consists of one old strand and one new strand. ||•|| nd All ↓ 2 round of replication. ||•|| ||•|| rd All ↓ 3 round of replication. ||•|| ||•|| ||•|| ||•|| ½ and ½ , or ½ like IV and ½ like III. Since DNA replication is known to be semiconservative, the results expected in the experiment are ½ like IV and ½ like III. ************************************************************************ Page 9 The following choices are used for questions 11 - 14. (a) (b) (c) (d) (e) Pribnow box CAAT box -35 consensus sequence Shine-Dalgarno sequence Hogness box d 11 A sequence in the leader region of mRNA, believed to be responsible for orienting the mRNA on the small ribosome subunit in the initiation of translation. a 12 In prokaryotes, a promoter consensus sequence 10 bases upstream from the first base transcribed. e 13 In eukaryotes, a promoter consensus sequence 25 bases upstream from the first base transcribed. Very few books use the term “Hogness box” anymore, using instead the term “TATA box” You will not use the term “Hogness box” on your exam. b 14 In eukaryotes, a promoter consensus sequence 70 bases upstream from the first base transcribed. ************************************************************************ 15 Rho factors (a) (b) (c) (d) (e) 16 are responsible for the termination of translation. are responsible for the termination of transcription in eukaryotes. are responsible for the termination of transcription in prokaryotes. are responsible for the initiation of transcription in eukaryotes. are responsible for the initiation of transcription in prokaryotes. Sigma factors (a) (b) (c) (d) (e) assist the small ribosome subunit in recognizing mRNA during the initiation of translation. assist prokaryotic RNA polymerase in recognizing the origin of replication during the initiation of replication. assist prokaryotic RNA polymerase in recognizing the promoter region during the initiation of transcription. assist eukaryotic RNA polymerase in recognizing the promoter region during the initiation of transcription. assist eukaryotic RNA polymerase in recognizing the origin of replication during the initiation of replication. Page 10 17 In post-transcriptional processing of eukaryotic mRNA, the enzyme terminal transferase is responsible for (a) (b) (c) (d) (e) The addition of a poly-A tail to the 3' end of the mRNA. The addition of a methylated guanosine cap to the 3' end of the mRNA. The formation of peptide bond between the amino group of one amino acid and the carboxyl group of another amino acid. The addition of a poly-A tail to the 5' end of the mRNA. The addition of a methylated guanosine cap to the 5' end of the mRNA. ************************************************************************ The nucleotide sequence listed below represents the transcriptional template strand of a gene. Template DNA strand 18 Which of the following is the nontemplate DNA strand complementary to the template strand? (a) (b) (c) (d) (e) 19 5' CTACGTAGTTGAAGACAT 3' 3' CTACGTAGTTGAAGACAT 5' 5' AUGUCUUCAACUACGUAG 3' 5' ATGTCTTCAACTACGTAG 3' 3' ATGTCTTCAACTACGTAG 5' Which of the following is the mRNA transcribed from the template DNA (assume that there is no intron splicing or other processing). (a) (b) (c) (d) (e) 20 3' TACAGAAGTTGATGCATC 5' 3' AUGUCUUCAACUACGUAG 5' 5' AUGUCUUCAACUACGUAG 3' 3' UACAGAAGUUGAUGCAUC 5' 5' UACAGAAGUUGAUGCAUC 3' 5' GAUUCUACUUCAGACGAU 3' Which of the following is the peptide that is produced when the mRNA is translated? (The standard 3-letter abbreviations for amino acids are used.) (a) (Amino end) Tyr Arg Ser (Carboxyl end) (b) (Amino end) Asp Ser Thr Ser Asp Asp (Carboxyl end) (c) (Carboxyl end) Asp Ser Thr Ser Asp Asp (Amino end) (d) (Carboxyl end) Met Ser Ser Thr Thr (Amino end) (e) (Amino end) Met Ser Ser Thr Thr (Carboxyl end) *********************************************************************** Page 11 21 Which of the following best describes the action of an aminoacyl tRNA synthetase enzyme? (a) (b) (c) (d) (e) 22` Which of the following is used directly as an energy source by enzymes in the initiation and elongation stages of translation? (a) (b) (c) (d) (e) 23 It attaches the amino end of an amino acid to the 5' end of a tRNA molecule. It attaches the carboxyl end of an amino acid to the 5' end of a tRNA molecule. It attaches the carboxyl end of an amino acid to the 3' end of a tRNA molecule. It attaches the amino end of an amino acid to the 3' end of a tRNA molecule. It attaches the anticodon of a tRNA to the codon on the mRNA. ATP CTP GTP UTP lactose A certain mutation in E. coli is a chain-termination (or nonsense) mutation. Compared to the wild type, one of the nucleotide bases in the mutant is substituted for another so that a sense codon UCG in the coding region is changed into the stop codon UAG. This means that translation terminates prematurely, resulting in a protein that is too short to carry out its enyzmatic functions. It is possible to isolate another E. coli mutant gene called a nonsense suppressor mutation. If cloned into the strain with the chain-termination mutation, the nonsense suppressor causes the normal wild phenotype to return to the strain. However, the nonsense suppressor gene does not encode the same protein as the chain termination gene. What does the nonsense suppressor encode? (a) (b) (c) (d) (e) The nonsense suppressor encodes a repressor protein that blocks transcription of the chain-termination gene. The nonsense suppressor encodes the aminoacyl tRNA synthetase specific for tRNA with the anticodon AGC. The nonsense suppressor encodes a tRNA molecule with the anticodon AGC. The nonsense suppressor encodes a tRNA molecule with the anticodon AUC. The nonsense suppressor encodes a Rho factor protein. Page 12 24 To which site on the ribosome does the next aminoacyl tRNA bind during the elongation phase of translation? (a) (b) (c) (d) (e) 1. The 5' site The 3' site The A site The P site The amino site What are the special features found at the 5' end and the 3' end of mature eukaryotic mRNA? (4 pt) At the 5’ end, there is a methylated guanosine cap attached via a 5’ -> 5’ triphophate linkage. It is added during posttranscriptional processing by a capping enzyme. At the 3’ end is a poly-A tail consisting of about 100 – 200 consecutive adenosine nucleotides. It is added during post-transcriptional processing by a terminal transferase enzyme. 2. The nucleotide sequence listed below represents the nontemplate strand of a gene. Please give: (a) the nucleotide sequence of the template strand, (b) the nucleotide sequence of the mRNA formed (assuming no introns are present), and (c) the amino acid sequence of the peptide chain encoded by this sequence. Be sure to include which ends are 5', 3', amino, or carboxyl. (16 pt) nontemplate DNA strand 5' CGATGTCTTCAACTACGTAG 3' template DNA strand: 3’GCTACAGAAGTTGATGCATC 5’ mRNA: 5’CGAUGUCUUCAACUACGUAG 3’ Peptide chain: (amino end) met - ser - ser - thr - thr (carboxyl end) (starting at an initiation codon) 3. Each of the following is related to or involved in DNA replication in the bacterium Escherichia coli. Please briefly describe or state the role of each. Note: for enzymes, you should state each separate enzymatic activity, and state its purpose. (2 pt each) (a) DNA ligase Forms the final phosphodoester bond between two fragments of a nascent chain during DNA replication. (b) Primase: Creates short RNA primers that are needed by DNA polymerase, because DNA polymerase cannot initiate the de novo synthesis of a new chain. (c) DNA polymerase I: This enzyme has 5’ -> 3’ polymerase activity capable of elongating a nascent strand. It also has 3’ exonuclease activity that serves to remove mismatched bases (proofreading function). It also has 5’ exonuclease activity that is responsible for primer excision. Its major role in vivo is primer excision and proofreading. Page 13 (d) DNA polymerase III: This enzyme has 5’-> 3’ poylmerase activity and also 3’ exonuclease (proofreading) activity, but no 5’ exonuclease activity. It is responsible for most of the nascent chain elongation during replication in E. coli. (e) Okasaki fragments: These are short fragments of nascent strand DNA representing the discontinuously synthesized strand (lagging strand). 6. In an experiment, cells of actively growing E. coli were incubated for several minutes with radiolabeled deoxyribonucleosides (dNTPs). The DNA was isolated from the cells, denatured into single strands, and separated according to size by gradient centrifugation. The radioactivity in the tube corresponded to a single longer piece of DNA and a heterogeneous mixture of much shorter DNA fragments. Briefly explain these results. In your answer, you should use big words like “discontinuous.” (4 pt) The longer strand represented continuous strand synthesis, and the shorter fragments represented discontinuous strand synthesis (Okasaki fragments).. 5. Eukaryotic cells can be grown in a medium containing bromodeoxyuridine (BrdU), which substitutes for thymidine during DNA replication. A fluorescent dye that binds to double-stranded DNA makes it possible to recognize chromosomes or chromatids with BrdU incorporated in their DNA. You can tell the difference between unlabeled chromatids, chromatids with one strand of the DNA double helix labeled, and chromatids with both strands of the DNA double helix labeled. The chromatid fluoresces brightly when only one strand contains BrdU, but weakly when both strands contain BrdU. The following diagrams represent the fluorescence of metaphase chromosomes. Page 14 A group of cultured eukaryotic cells, all at the G1 stage, were placed in a medium containing BrdU and allowed to grow. Samples were removed during the first, second, and third mitotic divisions. Mitosis in each of these samples was arrested at metaphase using colchicine. Chromosomes were isolated from cells in the samples and examined for fluorescence. What will be the appearance of the metaphase chromosomes isolated in the first, second, and third mitotic divisions after transferring the cells to the BrdU medium? Answer this question by sketching the chromosomes, as shown in the diagram. Use double hatch marks to indicate bright fluorescence, and single hatch marks for weak fluorescence. (6 pt) Here are two hints. Hint 1: Chromosomes isolated at metaphase are in the replicated state (a pair of chromatids connected at the centromere) Hint 2: Each chromatid in a replicated chromosome has one double helix. The two helices are the products of DNA replication. See pgs. 8 & 9 for an explanation. This question asks you to predict what the appearance should be, so you should state the results for SEMICONSERVATIVE replication. Page 15