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Chapter 17: Molecular Basis of Inheritance. Overview: The Flow of Genetic Information The information content of DNA is in the form of specific sequences.

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Presentation on theme: "Chapter 17: Molecular Basis of Inheritance. Overview: The Flow of Genetic Information The information content of DNA is in the form of specific sequences."— Presentation transcript:

1 Chapter 17: Molecular Basis of Inheritance

2 Overview: The Flow of Genetic Information The information content of DNA is in the form of specific sequences of nucleotides Determines proteins built! Gene expression, the process by which DNA directs protein synthesis, includes two stages: transcription and translation

3 Fig. 17-1

4 Transcription and Translation RNA is the intermediate between genes and the proteins for which they code Transcription is the synthesis of RNA under the direction of DNA (DNA written into RNA) Transcription produces messenger RNA (mRNA) which takes code to the ribosome Translation is the synthesis of a polypeptide, mRNA has the code to build the protein Ribosomes are the sites of translation

5 In prokaryotes, mRNA produced by transcription is immediately translated without more processing In a eukaryotic cell, the nuclear envelope separates transcription from translation How do prokaryotic and eukaryotic organisms differ?

6 Fig. 17-3 TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide (a) Bacterial cell Nuclear envelope TRANSCRIPTION RNA PROCESSING Pre-mRNA DNA mRNA TRANSLATION Ribosome Polypeptide (b) Eukaryotic cell

7 Codons: Triplets of Bases The flow of information from gene to protein is based on a triplet code: a series of three- nucleotide words or bases Each triplet codon CODES for an amino acid Example: AGT at a particular position on a DNA strand results in the placement of the amino acid serine at the corresponding position of the polypeptide to be produced

8 In transcription, one of the two DNA strands called the template strand provides a template for ordering the sequence of nucleotides in an RNA transcript The Template Strand

9 In RNA: -Pairs are similar as in DNA, BUT -There is NO Thymine, instead Uracil A T C G Synthesis of mRNA (messenger RNA strand)

10 During translation, the mRNA base triplets, called codons, are read in the 5’ to 3’ direction Each codon specifies the amino acid to be placed at the corresponding position along a polypeptide

11 Fig. 17-4 DNA molecule Gene 1 Gene 2 Gene 3 DNA template strand TRANSCRIPTION TRANSLATION mRNA Protein Codon Amino acid

12 Cracking the Code All 64 codons were deciphered by the mid- 1960s Of the 64 triplets, 61 code for amino acids; 3 triplets are “stop” signals to end translation The genetic code is redundant but no codon specifies more than one amino acid Codons must be read in the correct reading frame (correct groupings) in order for the correct amino acid to be added and specified polypeptide to be produced The fat cat in the hat

13 Fig. 17-5 Second mRNA base First mRNA base (5 end of codon) Third mRNA base (3 end of codon)

14 Evolution of the Genetic Code The genetic code is nearly universal, shared by the simplest bacteria to the most complex animals Genes can be transcribed and translated after being transplanted from one species to another Ex insulin production

15 Transcription in Detail Helicase unwinds the DNA, just like in replication RNA polymerase pries the DNA strands apart and hooks together the RNA nucleotides (A-U, C-G, T-A)

16 Promoter: The DNA sequence where RNA polymerase attaches in bacteria, the sequence signaling the end of transcription is called the terminator (different in eukaryotes) The stretch of DNA that is transcribed is called a transcription unit

17 Fig. 17-7 Promoter Transcription unit Start point DNA RNA polymerase 5 53 3 Initiation 1 2 3 5 5 3 3 Unwound DNA RNA transcript Template strand of DNA Elongation Rewound DNA 5 5 5 5 5 3 3 3 3 RNA transcript Termination 5 5 3 3 3 5 Completed RNA transcript Newly made RNA Template strand of DNA Direction of transcription (“downstream”) 3 end RNA polymerase RNA nucleotides Nontemplate strand of DNA Elongation

18 Synthesis of an RNA Transcript The three stages of transcription: –Initiation –Elongation –Termination

19 Fig. 17-8 A eukaryotic promoter includes a TATA box 3 1 2 3 Promoter TATA box Start point Template DNA strand 5 3 5 Transcription factors Several transcription factors must bind to the DNA before RNA polymerase II can do so. 5 53 3 Additional transcription factors bind to the DNA along with RNA polymerase II, forming the transcription initiation complex. RNA polymerase II Transcription factors 5 5 5 3 3 RNA transcript Transcription initiation complex

20 RNA Polymerase Binding and Initiation of Transcription Promoters signal the initiation of RNA synthesis Transcription factors (proteins) mediate the binding of RNA polymerase and the initiation of transcription RNA polymerase II pries DNA apart and adds RNA nucleotides TATA box is a promoter DNA sequence (tells transcription factors and RNA polymerase II where to bind) Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

21 Elongation of the RNA Strand As RNA polymerase moves along the DNA, it untwists the double helix, 10 to 20 bases at a time

22 Termination of Transcription The mechanisms of termination are different in bacteria and eukaryotes In bacteria, the polymerase stops transcription at the end of the terminator In eukaryotes, reaches a polyadenylation signal “AAUAA” and and associated proteins cut mRNA free, called “pre-mRNA” Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

23 Eukaryotic cells modify RNA after transcription Enzymes in the eukaryotic nucleus modify pre- mRNA before the genetic messages are dispatched to the cytoplasm During RNA processing, both ends of the primary transcript are usually altered Also, usually some interior parts of the molecule are cut out, and the other parts spliced together

24 Alteration of mRNA Ends Each end of a pre-mRNA molecule is modified in a particular way: –The 5’ end receives a modified Guanine nucleotide (5’ cap) –The 3’ end gets a poly-A tail (many A’s) These modifications share several functions: –They seem to facilitate the export of mRNA –They protect mRNA from hydrolytic enzymes –They help ribosomes attach to the 5’ end Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

25 Fig. 17-9 Protein-coding segment Polyadenylation signal 3’3’ 3’ UTR5’ UTR 5’5’ 5’ Cap Start codon Stop codon Poly-A tail G PPPAAUAAA AAA …

26 Split Genes and RNA Splicing Most eukaryotic genes and their RNA transcripts have long noncoding stretches of nucleotides that lie between coding regions These noncoding regions are called introns The other regions are called exons because they are eventually expressed, usually translated into amino acid sequences RNA splicing removes introns and joins exons, creating an mRNA molecule with a continuous coding sequence

27 Fig. 17-10 Pre-mRNA mRNA Coding segment Introns cut out and exons spliced together 5’ Cap Exon Intron 5’5’ 1 30 31104 ExonIntron 105 Exon 146 3’3’ Poly-A tail 5 Cap 5’ UTR3’ UTR 1 146

28 In some cases, RNA splicing is carried out by spliceosomes Spliceosomes consist of a variety of proteins and several small nuclear ribonucleoproteins (snRNPs) that recognize the splice sites, cut away introns

29 Fig. 17-11-3 RNA transcript (pre-mRNA) Exon 1Exon 2Intron Protein snRNA snRNPs Other proteins 5 5 Spliceosome components Cut-out intron mRNA Exon 1 Exon 2 5

30 The Functional and Evolutionary Importance of Introns Some genes can encode more than one kind of polypeptide, depending on which segments are treated as exons during RNA splicing Such variations are called alternative RNA splicing Because of alternative splicing, the number of different proteins an organism can produce is much greater than its number of genes

31 Molecular Components of Translation A cell translates an mRNA message into protein with the help of transfer RNA (tRNA) tRNA –Each carries a specific amino acid on one end –Each has an anticodon on the other end; the anticodon base-pairs with a complementary codon on mRNA http://www.youtube.com/watch? v=Ikq9AcBcohA

32 Fig. 17-13 Polypeptide Ribosome Amino acids tRNA with amino acid attached tRNA Anticodon Trp Phe Gly Codons 3 5 mRNA

33 Fig. 17-14 Amino acid attachment site 3 5 Hydrogen bonds Anticodon (a) Two-dimensional structure Amino acid attachment site 5 3 Hydrogen bonds 3 5 Anticodon (c) Symbol used in this book (b) Three-dimensional structure

34 Fig. 17-15-4 Amino acid Aminoacyl-tRNA synthetase (enzyme) ATP Adenosine PPP P P P i P P i i tRNA Aminoacyl-tRNA synthetase Computer model Adenosine P Aminoacyl-tRNA (“charged tRNA”)

35 Accurate translation requires two steps: –First: a correct match between a tRNA and an amino acid, done by the enzyme aminoacyl- tRNA synthetase –Second: a correct match between the tRNA anticodon and an mRNA codon Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

36 Ribosomes The two ribosomal subunits (large and small) are made of proteins and ribosomal RNA (rRNA)

37 Fig. 17-16 Growing polypeptide Exit tunnel Large subunit Small subunit tRNA molecules E P A mRNA 5 3 (a) Computer model of functioning ribosome P site (Peptidyl-tRNA binding site) E site (Exit site) A site (Aminoacyl- tRNA binding site) E P A Large subunit mRNA binding site Small subunit (b) Schematic model showing binding sites Amino end Growing polypeptide Next amino acid to be added to polypeptide chain mRNA tRNA E 3 5 Codons (c) Schematic model with mRNA and tRNA

38 A ribosome has three binding sites for tRNA: –The P site holds the tRNA that carries the growing polypeptide chain –The A site holds the tRNA that carries the next amino acid to be added to the chain –The E site is the exit site, where discharged tRNAs leave the ribosome

39 Building a Polypeptide The three stages of translation: –Initiation –Elongation –Termination All three stages require protein “factors” that aid in the translation process

40 Initiation of Translation The initiation stage of translation brings together mRNA, a tRNA with the first amino acid, and the two ribosomal subunits First, a small ribosomal subunit binds with mRNA and a special initiator tRNA Then the small subunit moves along the mRNA until it reaches the start codon (AUG) Proteins called initiation factors bring in the large subunit that completes the translation initiation complex Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

41 Fig. 17-17 3’3’ 3’3’ 5’5’ 5’5’ U A C G Met Initiator tRNA mRNA 5’5’ 3’3’ Start codon mRNA binding site Small ribosomal subunit 5’5’ P site Translation initiation complex 3’3’ EA Met Large ribosomal subunit

42 Elongation of the Polypeptide Chain During the elongation stage, amino acids are added one by one to the preceding amino acid Each addition involves proteins called elongation factors and occurs in three steps: codon recognition, peptide bond formation, and translocation Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

43 Fig. 17-18-1 Amino end of polypeptide mRNA 5 3 E P site A site

44 Fig. 17-18-2 Amino end of polypeptide mRNA 5 3 E P site A site GTP GDP E P A

45 Fig. 17-18-3 Amino end of polypeptide mRNA 5 3 E P site A site GTP GDP E P A E PA

46 Fig. 17-18-4 Amino end of polypeptide mRNA 5 3 E P site A site GTP GDP E P A E PA GTP Ribosome ready for next aminoacyl tRNA E P A

47 Termination of Translation Termination occurs when a stop codon in the mRNA reaches the A site of the ribosome The A site accepts a protein called a release factor The release factor causes the addition of a water molecule instead of an amino acid This reaction releases the polypeptide, and the translation assembly then comes apart

48 Fig. 17-19-1 Release factor 3 5 Stop codon (UAG, UAA, or UGA)

49 Fig. 17-19-2 Release factor 3 5 Stop codon (UAG, UAA, or UGA) 5 3 2 Free polypeptide 2 GDP GTP

50 Fig. 17-19-3 Release factor 3 5 Stop codon (UAG, UAA, or UGA) 5 3 2 Free polypeptide 2 GDP GTP 5 3

51 Completing and Targeting the Functional Protein Often translation is not sufficient to make a functional protein Polypeptide chains are modified after translation Completed proteins are targeted to specific sites in the cell Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

52 Protein Folding and Post- Translational Modifications During and after synthesis, a polypeptide chain spontaneously coils and folds into its three- dimensional shape Proteins may also require post-translational modifications before doing their job Some polypeptides are activated by enzymes that cleave them Other polypeptides come together to form the subunits of a protein

53 Targeting Polypeptides to Specific Locations Two populations of ribosomes are evident in cells: free ribsomes (in the cytosol) and bound ribosomes (attached to the ER) Free ribosomes mostly synthesize proteins that function in the cytosol Bound ribosomes make proteins of the endomembrane system and proteins that are secreted from the cell Ribosomes are identical and can switch from free to bound

54 Polypeptide synthesis always begins in the cytosol Synthesis finishes in the cytosol unless the polypeptide signals the ribosome to attach to the ER Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

55 Mutations Mutations are changes in the genetic material of a cell or virus Point mutations are changes in just one base pair of a gene The change of a single nucleotide in a DNA template strand can lead to the production of an abnormal protein

56 Fig. 17-22 Wild-type hemoglobin DNA mRNA Mutant hemoglobin DNA mRNA 3 3 3 3 3 3 5 5 5 5 5 5 CCTT T T G G A A A A AA A GG U Normal hemoglobinSickle-cell hemoglobin Glu Val

57 Types of Point Mutations Point mutations within a gene can be divided into two general categories –Base-pair substitutions –Base-pair insertions or deletions

58 Fig. 17-23 Wild-type 3DNA template strand 5 5 5 3 3 Stop Carboxyl end Amino end Protein mRNA 3 3 3 5 5 5 A instead of G U instead of C Silent (no effect on amino acid sequence) Stop T instead of C 3 3 3 5 5 5 A instead of G Stop Missense A instead of T U instead of A 3 3 3 5 5 5 Stop Nonsense No frameshift, but one amino acid missing (3 base-pair deletion) Frameshift causing extensive missense (1 base-pair deletion) Frameshift causing immediate nonsense (1 base-pair insertion) 5 5 5 3 3 3 Stop missing 3 3 3 5 5 5 Stop 5 5 5 3 3 3 Extra U Extra A (a) Base-pair substitution(b) Base-pair insertion or deletion

59 Substitutions A base-pair substitution replaces one nucleotide and its partner with another pair of nucleotides Silent mutations have no effect on the amino acid produced by a codon because of redundancy in the genetic code Missense mutations still code for an amino acid, but not necessarily the right amino acid Nonsense mutations change an amino acid codon into a stop codon, nearly always leading to a nonfunctional protein Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

60 Insertions and Deletions Insertions and deletions are additions or losses of nucleotide pairs in a gene These mutations have a disastrous effect on the resulting protein more often than substitutions do Insertion or deletion of nucleotides may alter the reading frame, producing a frameshift mutation Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

61 Mutagens Spontaneous mutations can occur during DNA replication, recombination, or repair Mutagens are physical or chemical agents that can cause mutations Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

62 Comparing Gene Expression in Bacteria, Archaea, and Eukarya Bacteria and eukarya differ in their RNA polymerases, termination of transcription and ribosomes; archaea tend to resemble eukarya in these respects Bacteria can simultaneously transcribe and translate the same gene In eukarya, transcription and translation are separated by the nuclear envelope In archaea, transcription and translation are likely coupled Copyright © 2008 Pearson Education Inc., publishing as Pearson Benjamin Cummings

63 Fig. 17-25 TRANSCRIPTION RNA PROCESSING DNA RNA transcript 3 5 RNA polymerase Poly-A RNA transcript (pre-mRNA) Intron Exon NUCLEUS Aminoacyl-tRNA synthetase AMINO ACID ACTIVATION Amino acid tRNA CYTOPLASM Poly-A Growing polypeptide 3 Activated amino acid mRNA TRANSLATION Cap Ribosomal subunits Cap 5 E P A A Anticodon Ribosome Codon E


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