From DNA to Protein. Chapter 14
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1 From DNA to Protein Chapter 14
2 Impacts, Issues: Ricin and your Ribosomes Ricin is toxic because it inactivates ribosomes, the organelles which assemble amino acids into proteins, critical to life processes
3 14.1 DNA, RNA, and Gene Expression What is genetic information and how does a cell use it?
4 The Nature of Genetic Information Each strand of DNA consists of a chain of four kinds of nucleotides: A, T, G and C The sequence of the four bases in the strand is the genetic information
5 Converting a Gene to an RNA Transcription Enzymes use the nucleotide sequence of a gene to synthesize a complementary strand of RNA DNA is transcribed to RNA Most RNA is single stranded RNA uses uracil in place of thymine RNA uses ribose in place of deoxyribose
6 Ribonucleotides and Nucleotides
7 Ribonucleotides and Nucleotides
8 base (guanine) 3 phosphate groups sugar (ribose) guanine G (RNA) guanosine triphosphate A Guanine, one of the four nucleotides in RNA. The others (adenine, uracil, and cytosine) differ only in their component bases. Three of the four bases in RNA nucleotides are identical to the bases in DNA nucleotides. Fig. 14-2a, p. 216
9 base (guanine) 3 phosphate groups sugar (deoxyribose) guanine G (DNA) deoxyguanosine triphosphate B Compare the DNA nucleotide guanine. The only structural difference between the RNA and DNA versions of guanine (or adenine, or cytosine) is the functional group on the 2 carbon of the sugar. Fig. 14-2b, p. 216
10 DNA and RNA
11 adenine A DNA RNA adenine A NH 2 deoxyribonucleic acid ribonucleic acid NH 2 N C C N N C C nucleotide N HC HC N C CH base N C CH N N sugar guanine G phosphate guanine G O backbone O N C C N C NH C NH HC HC N C N C N C C NH 2 N NH 2 cytosine C NH 2 cytosine C NH 2 HC C N HC C N HC N C O HC N C O thymine T O base pair uracil U O CH 3 C C NH HC C NH HC C O HC C O N N DNA has one function: It RNAs have various Nucleotide permanently stores a cell s functions. Some serve Nucleotide bases of DNA genetic information, which as disposable copies of bases of RNA is passed to offspring. DNA s genetic message; others are catalytic. Fig. 14-3, p. 217
12 RNA in Protein Synthesis Messenger RNA (mrna) Contains information transcribed from DNA Ribosomal RNA (rrna) Main component of ribosomes, where polypeptide chains are built Transfer RNA (trna) Delivers amino acids to ribosomes
13 Converting mrna to Protein Translation The information carried by mrna is decoded into a sequence of amino acids, resulting in a polypeptide chain that folds into a protein mrna is translated to protein rrna and trna translate the sequence of base triplets in mrna into a sequence of amino acids
14 Gene Expression A cell s DNA sequence (genes) contains all the information needed to make the molecules of life Gene expression A multistep process including transcription and translation, by which genetic information encoded by a gene is converted into a structural or functional part of a cell or body
15 14.1 Key Concepts DNA to RNA to Protein Proteins consist of polypeptide chains The chains are sequences of amino acids that correspond to sequences of nucleotide bases in DNA called genes The path leading from genes to proteins has two steps: transcription and translation
16 14.2 Transcription: DNA to RNA RNA polymerase assembles RNA by linking RNA nucleotides into a chain, in the order dictated by the base sequence of a gene A new RNA strand is complementary in sequence to the DNA strand from which it was transcribed
17 DNA Replication and Transcription DNA replication and transcription both synthesize new molecules by base-pairing In transcription, a strand of mrna is assembled on a DNA template using RNA nucleotides Uracil (U) nucleotides pair with A nucleotides RNA polymerase adds nucleotides to the transcript
18 Base-Pairing in DNA Synthesis and Transcription
19 DNA template New DNA strand DNA template RNA transcript Stepped Art Fig. 14-4, p. 218
20 The Process of Transcription RNA polymerase and regulatory proteins attach to a promoter (a specific binding site in DNA close to the start of a gene) RNA polymerase moves over the gene in a 5' to 3' direction, unwinds the DNA helix, reads the base sequence, and joins free RNA nucleotides into a complementary strand of mrna
21 Transcription
22 gene region newly forming RNA transcript RNA polymerase, the enzyme that catalyzes transcription DNA template winding up DNA template unwinding A RNA polymerase binds to a promoter in the DNA, along with regulatory proteins. The binding positions the polymerase near a gene in the DNA. In most cases, the nucleotide sequence of the gene occurs on only one of the two strands of DNA. Only the complementary strand will be translated into RNA. B The polymerase begins to move along the DNA and unwind it. As it does, it links RNA nucleotides into a strand of RNA in the order specified by the base sequence of the DNA. The DNA double helix winds up again after the polymerase passes. The structure of the opened DNA molecule at the transcription site is called a transcription bubble, after its appearance. Fig. 14-5a, p. 218
23 Fig. 14-5b, p. 219
24 transcription site 5 growing RNA transcript 3 C What happened in the gene region? RNA polymerase catalyzed the covalent bonding of many nucleotides to one another to form an RNA strand. The base sequence of the new RNA strand is complementary to the base sequence of its DNA template a copy of the gene. Fig. 14-5b, p. 219
25 Animation: Gene transcription details
26 Transcription Many RNA polymerases can transcribe a gene at the same time
27 RNA transcripts DNA molecule Fig. 14-6, p. 219
28 14.2 Key Concepts DNA to RNA: Transcription During transcription, one strand of a DNA double helix is a template for assembling a single, complementary strand of RNA (a transcript) Each transcript is an RNA copy of a gene
29 14.3 RNA and the Genetic Code Base triplets in an mrna are words in a proteinbuilding message Two other classes of RNA (rrna and trna) translate those words into a polypeptide chain
30 Post-Transcriptional Modifications In eukaryotes, RNA is modified before it leaves the nucleus as a mature mrna Introns Nucleotide sequences that are removed from a new RNA Exons Sequences that stay in the RNA
31 Alternative Splicing Alternative splicing Allows one gene to encode different proteins Some exons are removed from RNA and others are spliced together in various combinations After splicing, transcripts are finished with a modified guanine cap at the 5' end and a poly- A tail at the 3' end
32 Post-Transcriptional Modifications
33 gene DNA exon intron exon intron exon cap transcription into RNA poly-a tail RNA 5 3 snipped out snipped out mrna Fig. 14-7, p. 220
34 gene DNA exon intron exon intron exon cap transcription into RNA poly-a tail RNA 5 3 snipped out snipped out mrna Stepped Art Fig. 14-7, p. 220
35 Animation: Pre-mRNA transcript processing
36 mrna The Messenger mrna carries protein-building information to ribosomes and trna for translation Codon A sequence of three mrna nucleotides that codes for a specific amino acid The order of codons in mrna determines the order of amino acids in a polypeptide chain
37 Genetic Information From DNA to mrna to amino acid sequence
38 DNA mrna mrna amino acids proline glutamic acid threonine lysine Fig. 14-8, p. 220
39 Genetic Code Genetic code Consists of 64 mrna codons (triplets) Some amino acids can be coded by more than one codon Some codons signal the start or end of a gene AUG (methionine) is a start codon UAA, UAG, and UGA are stop codons
40 Codons of the Genetic Code
41 Animation: Genetic code
42 rrna and trna The Translators trnas deliver amino acids to ribosomes trna has an anticodon complementary to an mrna codon, and a binding site for the amino acid specified by that codon Ribosomes, which link amino acids into polypeptide chains, consist of two subunits of rrna and proteins
43 Ribosomes
44 tunnel large subunit small subunit intact ribosome Fig , p. 221
45 trna
46 Fig a, p. 221
47 anticodon trp amino acid attachment site Fig a, p. 221
48 Fig b, p. 221
49 14.3 Key Concepts RNA Messenger RNA carries DNA s protein-building instructions Its nucleotide sequence is read three bases at a time Sixty-four mrna base triplets codons represent the genetic code Two other types of RNA interact with mrna during translation of that code
50 14.4 Translation: RNA to Protein Translation converts genetic information carried by an mrna into a new polypeptide chain The order of the codons in the mrna determines the order of the amino acids in the polypeptide chain
51 Translation Translation occurs in the cytoplasm of cells Translation occurs in three stages Initiation Elongation Termination
52 Initiation An initiation complex is formed A small ribosomal subunit binds to mrna The anticodon of initiator trna base-pairs with the start codon (AUG) of mrna A large ribosomal subunit joins the small ribosomal subunit
53 Elongation The ribosome assembles a polypeptide chain as it moves along the mrna Initiator trna carries methionine, the first amino acid of the chain The ribosome joins each amino acid to the polypeptide chain with a peptide bond
54 Termination When the ribosome encounters a stop codon, polypeptide synthesis ends Release factors bind to the ribosome Enzymes detach the mrna and polypeptide chain from the ribosome
55 Polysomes Many ribosomes may simultaneously translate the same mrna, forming polysomes
56 polysome p. 222
57 Translation in Eukaryotes
58 Initiation A A mature mrna leaves the nucleus and enters cytoplasm, which has many free amino acids, trnas, and ribosomal subunits. An initiator trna binds to a small ribosomal subunit and the mrna. mrna initiator trna small ribosomal subunit large ribosomal subunit B A large ribosomal subunit joins, and the cluster is now called an initiation complex. Stepped Art Fig (a-b), p. 222
59 Translation in Eukaryotes
60 Fig c, p. 223
61 Elongation C An initiator trna carries the amino acid methionine, so the first amino acid of the new polypeptide chain will be methionine. A second trna binds the second codon of the mrna (here, that codon is GUG, so the trna that binds carries the amino acid valine). A peptide bond forms between the first two amino acids (here, methionine and valine). Fig c, p. 223
62 Fig d, p. 223
63 D The first trna is released and the ribosome moves to the next codon in the mrna. A third trna binds to the third codon of the mrna (here, that codon is UUA, so the trna carries the amino acid leucine). A peptide bond forms between the second and third amino acids (here, valine and leucine). Fig d, p. 223
64 Fig e, p. 223
65 E The second trna is released and the ribosome moves to the next codon. A fourth trna binds the fourth mrna codon (here, that codon is GGG, so the trna carries the amino acid glycine). A peptide bond forms between the third and fourth amino acids (here, leucine and glycine). Fig e, p. 223
66 Fig f, p. 223
67 Termination F Steps d and e are repeated over and over until the ribosome encounters a stop codon in the mrna. The mrna transcript and the new polypeptide chain are released from the ribosome. The two ribosomal subunits separate from each other. Translation is now complete. Either the chain will join the pool of proteins in the cytoplasm or it will enter rough ER of the endomembrane system (Section 4.9). Fig f, p. 223
68 Animation: Translation
69 14.4 Key Concepts RNA to Protein: Translation Translation is an energy-intensive process by which a sequence of codons in mrna is converted to a sequence of amino acids in a polypeptide chain
70 14.5 Mutated Genes and Their Protein Products If the nucleotide sequence of a gene changes, it may result in an altered gene product, with harmful effects Mutations Small-scale changes in the nucleotide sequence of a cell s DNA that alter the genetic code
71 Common Mutations Base-pair-substitution May result in a premature stop codon or a different amino acid in a protein product Example: sickle-cell anemia Deletion or insertion Can cause the reading frame of mrna codons to shift, changing the genetic message Example: Huntington s disease
72 Common Mutations
73 part of DNA mrna transcribed from DNA resulting amino acid sequence base substitution in DNA altered mrna altered amino acid sequence deletion in DNA altered mrna altered amino acid sequence THREONINE PROLINE GLUTAMATE GLUTAMATE LYSINE THREONINE PROLINE VALINE GLUTAMATE LYSINE THREONINE PROLINE GLYCINE ARGININE A Part of the DNA, mrna, and amino acid sequence of the beta chain of a normal hemoglobin molecule. B A base-pair substitution in DNA replaces a thymine with an adenine. When the altered mrna is translated, valine replaces glutamate as the sixth amino acid of the new polypeptide chain. Hemoglobin with this chain is HbS sickle hemoglobin (Section 3.6). C Deletion of the same thymine causes a frameshift. The reading frame for the rest of the mrna shifts, and a different protein product forms. This mutation results in a defective hemoglobin molecule. The outcome is thalassemia, a type of anemia. Fig , p. 224
74 part of DNA mrna transcribed from DNA resulting amino acid sequence base substitution in DNA altered mrna altered amino acid sequence deletion in DNA altered mrna altered amino acid sequence THREONINE PROLINE GLUTAMATE GLUTAMATE LYSINE THREONINE PROLINE VALINE GLUTAMATE LYSINE THREONINE PROLINE GLYCINE ARGININE A Part of the DNA, mrna, and amino acid sequence of the beta chain of a normal hemoglobin molecule. B A base-pair substitution in DNA replaces a thymine with an adenine. When the altered mrna is translated, valine replaces glutamate as the sixth amino acid of the new polypeptide chain. Hemoglobin with this chain is HbS sickle hemoglobin (Section 3.6). C Deletion of the same thymine causes a frameshift. The reading frame for the rest of the mrna shifts, and a different protein product forms. This mutation results in a defective hemoglobin molecule. The outcome is thalassemia, a type of anemia. Stepped Art Fig , p. 224
75 Animation: Base-pair substitution
76 Animation: Frameshift mutation
77 What Causes Mutations? Transposable elements Segments of DNA that can insert themselves anywhere in a chromosomes Spontaneous mutations Uncorrected errors in DNA replication Harmful environmental agents Ionizing radiation, UV radiation, chemicals
78 McClintock s Transposable Elements
79 Mutations Caused by Radiation Ionizing radiation damages chromosomes, nonionizing (UV) radiation forms thymine dimers
80 Inherited Mutations Mutations in somatic cells of sexually reproducing species are not inherited Mutations in a germ cell or gamete may be inherited, with evolutionary consequences
81 14.5 Key Concepts Mutations Small-scale, permanent changes in the nucleotide sequence of DNA may result from replication errors, the activity of transposable elements, or exposure to environmental hazards Such mutation can change a gene s product
82 Summary: Protein Synthesis in Eukaryotic Cells
83 Transcription Assembly of RNA on unwound regions of DNA molecule mrna processing mrna rrna trna proteins mature mrna ribosomal transcripts subunits mature trna Translation At an intact ribosome, synthesis of a polypeptide chain at the binding sites for mrna and trnas Convergence of RNAs cytoplasmic pools of amino acids, ribosomal subunits, and trnas Protein Fig , p. 226
84 Animation: Protein synthesis summary
85 Animation: Structure of a ribosome
86 Animation: Structure of a trna
87 between prokaryotic and eukaryotic protein synthesis
88 Animation: Transcription
89 Animation: Uracil-thymine comparison
90 Video: Ricin and your ribosomes
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