1. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Chapter 17 From Gene to Protein

2. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Overview: The Flow of Genetic Information The information content of DNA – Is in the form of specific sequences of along the DNA strands

3. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The DNA inherited by an organism – Leads to specific traits by dictating the synthesis of proteins The process by which DNA directs protein synthesis, gene expression – Includes two stages, called

4. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Concept 17.1: Genes: – specify proteins via transcription and translation – Specify different types of RNA via transcription

5. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Basic Principles of Transcription and Translation Transcription – Is the synthesis of RNA under the direction of DNA. It produces: messenger RNA (mRNA) ribosomal RNA (rRNA) Transfer RNA (tRNA)

6. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Translation – Is the actual synthesis of a polypeptide, which occurs under the direction of – Occurs on

7. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings In prokaryotes Transcription and translation occur simultaneously Figure 17.3a Prokaryotic cell. In a cell lacking a nucleus, mRNA produced by transcription is immediately translated without additional processing. (a) TRANSLATION TRANSCRIPTION DNA mRNA Ribosome Polypeptide

8. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings In eukaryotes Transcription and translation occur at different times RNA transcripts are modified before becoming true mRNA Figure 17.3b Eukaryotic cell. The nucleus provides a separate compartment for transcription. The original RNA transcript, called pre-mRNA, is processed in various ways before leaving the nucleus as mRNA. (b) TRANSCRIPTION RNA PROCESSING TRANSLATION mRNA DNA Pre-mRNA Polypeptide Ribosome Nuclear envelope

9. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cells are governed by a cellular chain of command – DNA  mRNA  protein Or – DNA  rRNA Or DNA  tRNA

10. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Genetic Code Genetic information – Is encoded as a sequence of base triplets, called

11. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings During transcription The DNA base sequence of a gene determines the sequence of bases of an mRNA molecule DNA molecule Gene 1 Gene 2 Gene 3 DNA strand (template) TRANSCRIPTION mRNA Protein TRANSLATION Amino acid ACC AAACCGAG T UGG U UU G GC UC A Trp Phe Gly Ser Codon 3 5 3 5 Figure 17.4

12. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Genetic Code A codon in messenger RNA – Is either translated into an amino acid or serves as a translational stop signal Figure 17.5 Second mRNA base UCA G U C A G UUU UUC UUA UUG CUU CUC CUA CUG AUU AUC AUA AUG GUU GUC GUA GUG Met or start Phe Leu lle Val UCU UCC UCA UCG CCU CCC CCA CCG ACU ACC ACA ACG GCU GCC GCA GCG Ser Pro Thr Ala UAU UAC UGU UGC TyrCys CAU CAC CAA CAG CGU CGC CGA CGG AAU AAC AAA AAG AGU AGC AGA AGG GAU GAC GAA GAG GGU GGC GGA GGG UGG UAA UAG Stop UGA Stop Trp His Gln Asn Lys Asp Arg Ser Arg Gly U C A G U C A G U C A G U C A G First mRNA base (5 end) Third mRNA base (3 end) Glu

13. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Evolution of the Genetic Code The genetic code is nearly universal – Shared by organisms from the simplest bacteria to the most complex animals

14. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Concept 17.2: Transcription RNA synthesis – Is catalyzed by which opens the DNA strands and connects the RNA nucleotides – Follows the same rules as DNA, except that in RNA, uracil substitutes for thymine

15. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Synthesis of an RNA Transcript The stages of transcription are – Initiation – Elongation – Termination Figure 17.7 Promoter Transcription unit RNA polymerase Start point 5353 3535 3535 5353 5353 3535 5353 3535 5 5 Rewound RNA transcript 3 3 Completed RNA transcript Unwound DNA RNA transcript Template strand of DNA DNA 1 Initiation. After RNA polymerase binds to the promoter, the DNA strands unwind, and the polymerase initiates RNA synthesis at the start point on the template strand. 2 Elongation. The polymerase moves downstream, unwinding the DNA and elongating the RNA transcript 5  3. In the wake of transcription, the DNA strands re-form a double helix. 3 Termination. Eventually, the RNA transcript is released, and the polymerase detaches from the DNA.

16. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings RNA Polymerase Binding and Initiation of Transcription signal the initiation site for transcription – Help eukaryotic RNA polymerase recognize promoter sequences Figure 17.8 TRANSCRIPTION RNA PROCESSING TRANSLATION DNA Pre-mRNA mRNA Ribosome Polypeptide TATAA A A ATATTTT TATA box Start point Template DNA strand 5 3 3 5 Transcription factors 5 3 3 5 Promoter 5 3 3 5 5 RNA polymerase II Transcription factors RNA transcript Transcription initiation complex Eukaryotic promoters 1 Several transcription factors 2 Additional transcription factors 3

17. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Elongation of the RNA Strand As RNA polymerase moves along the DNA – It continues to untwist the double helix, adding successive complementary nucleotides, and extending the RNA molecule

18. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Elongation RNA polymerase Non-template strand of DNA RNA nucleotides 3 end C A U C C A A U T A G G T T A A C G U A T C A T CCA A T T G G 3 5 5 Newly made RNA Direction of transcription (“downstream”) Template strand of DNA

19. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Concept 17.3: Enzymes in the eukaryotic nucleus – Modify in specific ways before the genetic messages are dispatched to the cytoplasm

20. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Alteration of mRNA Ends Each end of a pre-mRNA molecule is modified in a particular way Figure 17.9 A modified guanine nucleotide added to the 5 end 50 to 250 adenine nucleotides added to the 3 end Protein-coding segment Polyadenylation signal Poly-A tail 3 UTR Stop codonStart codon 5 Cap 5 UTR AAUAAA AAA…AAA TRANSCRIPTION RNA PROCESSING DNA Pre-mRNA mRNA TRANSLATION Ribosome Polypeptide G P PP 5 3

21. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Split Genes and RNA Splicing RNA splicing – Removes introns and joins exons Figure 17.10 TRANSCRIPTION RNA PROCESSING DNA Pre-mRNA mRNA TRANSLATION Ribosome Polypeptide 5 Cap Exon Intron 1 5 30 31 Exon Intron 104105146 Exon 3 Poly-A tail Introns cut out and exons spliced together Coding segment 5 Cap 1 146 3 UTR Pre-mRNA mRNA

22. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Functional Importance of Introns The presence of introns – Allows for alternative RNA splicing

23. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Proteins often consist of discrete structural and functional regions called domains In many cases different code for the different domains in a protein Figure 17.12 Gene DNA Exon 1 Intron Exon 2IntronExon 3 Transcription RNA processing Translation Domain 3 Domain 1 Domain 2 Polypeptide

24. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Concept 17.4: Translation is the mRNA-directed synthesis of a polypeptide A cell translates an mRNA message into protein – With the help of

25. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Translation Figure 17.13 TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide Amino acids tRNA with amino acid attached Ribosome tRNA Anticodon mRNA Trp Phe Gly A G C A AA C C G U G GUUU GG C Codons 5 3

26. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Molecules of tRNA are not all identical – Each carries a specific on one end – Each has an on the other end

27. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Structure and Function of Transfer RNA A C C A tRNA molecule 3 C C A C G C U U A A G ACA C C U * G C * * G UGU * C U * GA G G U * * A * A A G U C A G A C C * C GA G A G G G * * G A C U C * A U U U A G G C G 5 Amino acid attachment site Hydrogen bonds Anticodon A Figure 17.14a

28. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ribosomes Facilitate the specific coupling of tRNA anticodons with mRNA codons during protein synthesis

29. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The ribosomal subunits Are constructed of proteins and RNA molecules named ribosomal RNA (rRNA) TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide Exit tunnel Growing polypeptide tRNA molecules E P A Large subunit Small subunit mRNA (a) 5 3 Figure 17.16a

30. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The ribosome has three binding sites for tRNA The P site The A site The E site Figure 17.16b EPA P site (Peptidyl-tRNA binding site) E site (Exit site) mRNA binding site A site (Aminoacyl- tRNA binding site) Large subunit Small subunit Schematic model showing binding sites. A ribosome has an mRNA binding site and three tRNA binding sites, known as the A, P, and E sites. This schematic ribosome will appear in later diagrams. (b)

31. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Figure 17.16c Amino end Growing polypeptide Next amino acid to be added to polypeptide chain tRNA mRNA Codons 3 5 Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon base-pairs with an mRNA codon. The P site holds the tRNA attached to the growing polypeptide. The A site holds the tRNA carrying the next amino acid to be added to the polypeptide chain. Discharged tRNA leaves via the E site. (c)

32. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Building a Polypeptide We can divide translation into three stages – Initiation – Elongation – Termination

33. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ribosome Association and Initiation of Translation The initiation stage of translation – Brings together mRNA, tRNA bearing the first amino acid of the polypeptide, and two subunits of a ribosome Large ribosomal subunit 2 Initiator tRNA mRNA mRNA binding site Small ribosomal subunit Translation initiation complex P site GDP GTP Start codon 1 Met U A C A U G EA 3 5 5 3 3 5 3 5 Figure 17.17

34. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Elongation of the Polypeptide Chain In the elongation stage of translation – Amino acids are added one by one to the preceding amino acid Figure 17.18 Amino end of polypeptide mRNA Ribosome ready for next aminoacyl tRNA E PA E P A E P A E P A GDP GTP GDP 2 2 site 5 3 TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide Codon recognition. The anticodon of an incoming tRNA base-pairs with the complementary mRNA codon in the A site. 1 Peptide bond formation. This step attaches the polypeptide to the tRNA in the A site. 2 Translocation. The ribosome translocates the tRNA in the A site to the P site. The empty tRNA in the P site is moved to the E site, where it is released. The mRNA moves along with its bound tRNAs, bringing the next codon to be translated into the A site. 3

35. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Termination of Translation The final stage of translation is termination when the ribosome reaches a in the mRNA Figure 17.19 Release factor Free polypeptide Stop codon (UAG, UAA, or UGA) 5 3 3 5 3 5 When a ribosome reaches a stop codon on mRNA, the A site of the ribosome accepts a protein called a release factor instead of tRNA. 1 The release factor hydrolyzes the bond between the tRNA in the P site and the last amino acid of the polypeptide chain. The polypeptide is thus freed from the ribosome. 23 The two ribosomal subunits and the other components of the assembly dissociate.

36. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Completing and Targeting the Functional Protein Polypeptide chains – Undergo modifications after the translation process Proteins may be modified in ways that affect their three-dimensional shape Other molecules may be attached to proteins, such as sugars, lipids, phosphates

37. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Targeting Polypeptides to Specific Locations Two populations of ribosomes are evident in cells – Free in the cytosol, and bound to ER Free ribosomes in the cytosol –

38. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Proteins destined for the endomembrane system or for secretion – Must be transported into the ER – Have signal peptides to which a signal- recognition particle (SRP) binds, enabling the translation ribosome to bind to the ER

39. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Figure 17.21 Ribosome mRNA Signal peptide Signal- recognition particle (SRP) SRP receptor protein Translocation complex CYTOSOL Signal peptide removed ER membrane Protein ERLUMEN The signal mechanism for targeting proteins to the ER Polypeptide synthesis begins on a free ribosome in the cytosol. 1 An SRP binds to the signal peptide. 2 The SRP binds to a receptor protein in the ER membrane. 3 The polypeptide begins translocating across the membrane. 4 A signal- cleaving enzyme cuts off the signal peptide. 5 The rest of the completed polypeptide leaves the ribosome and folds into its final conformation. 6

40. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings What is a gene? A gene – Is a region of DNA whose final product is either a or an

41. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings A summary of transcription and translation in a eukaryotic cell Figure 17.26 TRANSCRIPTION RNA is transcribed from a DNA template. DNA RNA polymerase RNA transcript RNA PROCESSING In eukaryotes, the RNA transcript (pre- mRNA) is spliced and modified to produce mRNA, which moves from the nucleus to the cytoplasm. Exon Poly-A RNA transcript (pre-mRNA) Intron NUCLEUS Cap FORMATION OF INITIATION COMPLEX After leaving the nucleus, mRNA attaches to the ribosome. CYTOPLASM mRNA Poly-A Growing polypeptide Ribosomal subunits Cap Aminoacyl-tRNA synthetase Amino acid tRNA AMINO ACID ACTIVATION Each amino acid attaches to its proper tRNA with the help of a specific enzyme and ATP. Activated amino acid TRANSLATION A succession of tRNAs add their amino acids to the polypeptide chain as the mRNA is moved through the ribosome one codon at a time. (When completed, the polypeptide is released from the ribosome.) Anticodon A C C AAA U G G U U UA U G U A C E A Ribosome 1 Poly-A 5 5 3 Codon 2 3 4 5