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Transcription:

DNA Introduction

Do you think DNA is important? T.V shows Movies Biotech Films News Cloning Genetic Engineering

At the most basic level DNA is a set of instructions for protein construction. Structural Enzymes Hormones

DNA Deoxyribonucleic acid Every living thing has DNA That means that you have something in common with a zebra, a tree, a mushroom and a beetle!!!! 5

What do you need to know? Structure of DNA How DNA replicates (reproduces)? How DNA transcribes RNA? How Ribosomes use the RNA to correctly assemble proteins?

Nobel Peace Prize The Double Helix, 1953 Outlined the Structure of the DNA Discovered the locking of bases Watson and Crick

DNA: video play in VLC media player 17:06-23:00

DNA as Genetic Material Macromolecules 1. Carbohydrate 2. Lipids 3. Proteins 4. Nucleic Acids (DNA an d RNA) -- Nucleic Acids are made up of nucleotides -- Nucleotides = monomers; nucleic acid = polymers --So, DNA and RNA are nucleic acids because they re made up of nucleotides Deoxyribonucleic acid Ribonucleic acid

DNA DNA contains genes, sequences of nucleotide bases These genes code for polypeptides (proteins) Proteins are used to build cells and do much of the work inside cells 10

Genes & Proteins Proteins are made of amino acids linked together by peptide bonds 20 different amino acids exist 11

Polypeptides Amino acid chains are called polypeptides 12

DNA Structure DNA is a polymer Each nucleotide consists of: 5 carbon sugar (deoxyribose) Nitrogen containing base attached to the sugar Phosphate group There are 4 different types of nucleotides found in DNA, differing only in the nitrogenous base 13

DNA Structure The 4 nucleotides are given one letter abbreviations as shorthard for the 4 bases A is for adenine G is for guanine C is for cytosine T is for thymine Purine Pyrimidine 14

Deciphering DNA's structure. # of purine bases = # of pyrimidine bases # of adenine bases = # of bases # of guanine bases = # of bases

DNA Structure Sugar-phosphate backbone Hydrogen bonds Hold DNA strands together Can only form between certain bases A & T 2 H-bonds G & C 3 H-bonds So, bases on one strand determines the bases on the other strand Paired bases = complementary base pairs DNA Double helix; long zipper, twisted into a coil

DNA: structure video play in VLC media player

DNA Begins the Process (review material) DNA is found inside the nucleus Proteins, however, are made in the cytoplasm of cells by organelles called ribosomes 22

Starting with DNA DNA s code must be copied and taken to the cytoplasm In the cytoplasm, this code must be read so amino acids can be assembled to make polypeptides (proteins) This process is called PROTEIN SYNTHESIS 23

COSMOS video: DVD (5minutes)

DNA Replication DNA Replication Website Produces 2 exact copies of DNA Occurs just prior to cell division

Steps of DNA Replication H bonds break between the two strands 2 strands of DNA molecules separate (enzyme) bases of free nucleotides in nucleus of cell fasten onto complementary bases on each exposed strand (enzymes to proofread) nucleotides join together making complete complementary strands 2 double stranded molecules of DNA exactly like the original are made (each with one old strand & one new strand) Replication forks replication begins at the same time at many different points along the DNA molecule enzymes link small segments into one long strand replication occurs quickly

Practice: DNA REPLICATION 5 CTG ACT CCT GAG GAG 3 3 5

Practice: DNA REPLICATION 5 CTG ACT CCT GAG GAG 3 3 GAC TGA GGA CTC CTC 5

DNA Replication http://www.youtube.com/watch?v=hyi2 myfbbxk

DNA: replication video play in VLC media player

PROTEIN SYNTHESIS

Protein Synthesis The production (synthesis) of proteins. 3 phases: 1. Transcription 2. RNA processing 3. Translation Remember: DNA RNA Protein

Pathway to Making a Protein DNA mrna trna (ribosomes) Protein 35

DNA RNA Protein DNA Nuclear membrane Transcription Eukaryotic Cell RNA Processing Translation mrna Ribosome Protein

Question: How does RNA (ribonucleic acid) differ from DNA (deoxyribonucleic acid)?

RNA differs from DNA 1. RNA has a sugar ribose DNA has a sugar deoxyribose 2. RNA contains uracil (U) DNA has thymine (T) 3. RNA molecule is single-stranded DNA is double-stranded 4. RNA molecule leaves the nucleus DNA stays in the nucleus 5. RNA is like a page/recipe DNA is like the whole book

1. Transcription DNA Nuclear membrane Eukaryotic Cell Transcription RNA Processing mrna Translation Ribosome Protein

1. Transcription The transfer of information in the nucleus from a DNA molecule to an RNA molecule. Only 1 DNA strand serves as the template When complete, RNA molecule can be released to the ribosomes. Transcription animation http://www.stolaf.edu/people/giannini/flashanimat/molgene tics/transcription.swf

Question: What is the enzyme responsible for the production of the RNA molecule?

Answer: RNA Polymerase Separates the DNA molecule by breaking the H- bonds between the bases. (Unzips the DNA) Then moves along one of the DNA strands and lines up complement RNA nucleotides with DNA. Bonds the sugar and phosphate groups of the RNA together. Does not bond the nitrogenous bases. RNA leaves the area between the DNA strands. DNA zips back up. RNA is processed and leaves the nucleus.

1. Transcription DNA RNA Polymerase mrna in construction

Question: What would be the complementary RNA strand for the following DNA sequence? DNA GCGTATG

Answer: DNA GCGTATG RNA CGCAUAC

Practice: DNA REPLICATION: 5 CTG ACT CCT GAG GAG 3 3 GAC TGA GGA CTC CTC 5 DNA to RNA (TRANSCRIPTION): 5 3

Practice: DNA REPLICATION: 5 CTG ACT CCT GAG GAG 3 3 GAC TGA GGA CTC CTC 5 DNA to RNA (TRANSCRIPTION): 5 GAC UGA GGA CUC CUC 3

DNA: transcription video play in VLC media player DNA: video clip 23:20-26:15

Types of RNA Types of RNA: A. messenger RNA (mrna) B. transfer RNA (trna) Remember: all produced in the nucleus!

A. Messenger RNA (mrna) Carries the information for a specific protein. Made up of 500 to 1000 nucleotides long. Made up of codons (sequence of three bases: AUG - methionine). Each codon, is specific for an amino acid.

A. Messenger RNA (mrna) mrna start codon A U G G G C U C C A U C G G C G C A U A A codon 1 codon 2 codon 3 codon 4 codon 5 codon 6 codon 7 protein methionine glycine serine isoleucine glycine alanine stop codon Primary structure of a protein aa1 aa2 aa3 aa4 aa5 aa6 peptide bonds

B. Transfer RNA (trna) Made up of 75 to 80 nucleotides long. Picks up the appropriate amino acid floating in the cytoplasm Transports amino acids to the mrna. Have anticodons that are opposites to mrna codons. Recognizes the appropriate codons on the mrna and bonds temporarily to them

B. Transfer RNA (trna) amino acid attachment site methionine amino acid U A C anticodon

Codons and Anticodons The 3 bases of an anticodon are complementary to the 3 bases of a codon Example: Codon ACU Anticodon UGA UGA ACU

The Genetic Code A codon designates an amino acid An amino acid may have more than one codon There are 20 amino acids, but 64 possible codons Some codons tell the ribosome to stop translating

The Genetic Code Use the code by reading from the center to the outside Example: AUG codes for Methionine

Name the Amino Acids GGG? UCA? CAU? GCA? AAA?

2. RNA Processing DNA Nuclear membrane Eukaryotic Cell Transcription RNA Processing mrna Translation Ribosome Protein

mrna Processing After the DNA is transcribed into RNA, editing must be done to the nucleotide chain to make the RNA functional Introns, non-functional segments of DNA are snipped out of the chain

mrna Editing Exons, segments of DNA that code for proteins, are then rejoined by the enzyme (ligase) A guanine triphosphate cap is added to the 5 end of the newly copied mrna A poly A tail is added to the 3 end of the RNA The newly processed mrna can then leave the nucleus

Result of Transcription CAP New Transcript Tail

DNA: video play in VLC media player 26:15-29:14) stop at restriction enzymes

3. Translation DNA Nuclear membrane Eukaryotic Cell Transcription RNA Processing mrna Translation Ribosome Protein

3. Translation Synthesis of proteins in the cytoplasm Involves the following: 1. mrna (codons) 2. trna (anticodons) 3. ribosomes 4. amino acids

3. Translation Three parts: 1. initiation: start codon (AUG) 2. elongation: building the polypeptide chain 3. termination: stop codon (UAG) Let s make a PROTEIN!!!!. Translation animation http://www.stolaf.edu/people/giannini/flashanimat/molgenetics/translation.swf

3. Translation Translation is the process of decoding the mrna into a polypeptide chain Ribosomes read mrna three bases or 1 codon at a time and construct the proteins 67

3. Translation Large subunit P Site A Site Small subunit A U G 68 mrna C U A C U U C G

Step 1- Initiation mrna transcript start codon AUG attaches to the small ribosomal subunit Small subunit attaches to large ribosomal subunit mrna transcript 69

Step 2 - Elongation As ribosome moves, two trna with their amino acids move into site A and P of the ribosome Peptide bonds join the amino acids 70

Initiation aa1 aa2 anticodon hydrogen bonds 1-tRNA U A C A U G codon 2-tRNA G A U C U A C U U C G A mrna

Elongation aa1 peptide bond aa2 aa3 3-tRNA anticodon hydrogen bonds 1-tRNA U A C A U G codon 2-tRNA G A A G A U C U A C U U C G A mrna

aa1 aa2 peptide bond aa3 1-tRNA U A C (leaves) 2-tRNA 3-tRNA G A A G A U A U G C U A C U U C G A Ribosomes move over one codon mrna

aa1 peptide bonds aa4 aa2 aa3 4-tRNA 2-tRNA 3-tRNA G C U A U G G A U G A A C U A C U U C G A A C U mrna

aa1 peptide bonds aa2 aa3 aa4 G 2-tRNA A U (leaves) 3-tRNA 4-tRNA G C U G A A A U G C U A C U U C G A A C U Ribosomes move over one codon mrna

aa1 aa2 peptide bonds aa3 aa4 aa5 5-tRNA U G A 3-tRNA 4-tRNA G A A G C U G C U A C U U C G A A C U mrna

aa1 aa2 peptide bonds aa3 aa4 aa5 5-tRNA 3-tRNA G A A 4-tRNA G C U G C U A C U U C G A A C U Ribosomes move over one codon U G A mrna

aa4 aa5 aa199 Termination aa3 aa2 primary structure of a protein aa200 aa1 A C U mrna 200-tRNA C A U G U U terminator or stop codon U A G

http://learn.genetics.utah.edu/content/begin/dna/transcribe/ End Product The end products of protein synthesis are the primary structure of a protein. A sequence of amino acid bonded together by peptide bonds. aa2 aa3 aa4 aa5 aa199 aa1 aa200

Question: The anticodon UAC belongs to a trna that recognizes and binds to a particular amino acid. What would be the DNA base code for this amino acid?

Answer: trna mrna DNA - UAC (anticodon) - AUG (codon) - TAC

Polyribosome Groups of ribosomes reading same mrna simultaneously producing many proteins (polypeptides). incoming large subunit 1 2 3 4 5 6 7 mrna incoming small subunit polypeptide

http://learn.genetics.utah.edu/co ntent/molecules/transcribe/ http://www.stolaf.edu/people/gia nnini/flashanimat/molgenetics/tr anscription.swf http://www.stolaf.edu/people/gia nnini/flashanimat/molgenetics/tra nslation.swf unzip a simplified dna, showing bases show pool of free nucleotides fill in complementary nucleotides sugars/phosphates bond end with 2 identical dna strands

http://learn.genetics.utah.edu/co ntent/molecules/transcribe/ http://www.stolaf.edu/people/gia nnini/flashanimat/molgenetics/tr anscription.swf http://www.stolaf.edu/people/gia nnini/flashanimat/molgenetics/tra nslation.swf unzip a simplified dna, showing bases show pool of free nucleotides fill in complementary nucleotides sugars/phosphates bond end with 2 identical dna strands