DNA Structure & Function (Outline)
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1 DNA Structure & Function (Outline) 1. Historical perspective (DNA as the genetic material): Genetic transformation and DNA DNA is the genetic material in bacterial viruses (phage) The base-pairing rule DNA structure 2. Basis for polarity of SS DNA and anti-parallel complementary strands of DNA 3. DNA replication models 4. Mechanism of DNA replication: steps and molecular machinery 5. Replication and the end of linear chromosomes- Molecular basis for aging 6. Fidelity of DNA replication
2 By definition, the genetic material of must be replicated DNA Replication direct the cell functions by providing information for production of proteins Flow of the genetic information (Gene Expression)
3 Current Connections to DNA structure and replication Why are we mortal with a limited life span?
4 DNA as the Genetic Material Time-line 1850 s Mendel Microsocopy: Mitosis and Meiosis 1902 Chromosome basis of inheritance (Thomas H. Morgan) 20 th century Work with bacteria and viruses
5 1928 Fredrick Griffith Experiments Concept of transformation (using Bacteria that cause pneumonia in mammals) 1944 Avery, McCarty, and MacLeod The transforming material is DNA DNA is the genetic material 1952 Hershey and Chase DNA is the genetic material in viruses that infect bacteria
6 Griffith- Phenomenon of Transformation, a change in genotype (genetic makeup) by a foreign substance that changes the phenotype (observed properties) of the cell
7 History of DNA Avery, MacLeod, and McCarty, DNA is the transforming material (Can convert Type R bacteria into S)
8 A phage, is a virus that infects bacteria and is made of DNA and protein. Alfred Hershey and Martha Chase- the genetic material of the phage T2 is DNA. mations.html#
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11 DNA Structure Prior to the 1950s, DNA is a polymer of nucleotides consisting of: a nitrogenous base a sugar a phosphate group
12 Polarity and antiparallel nature of the two DNA strands (5 and 3 ends)
13 Biochemical analysis of DNA: Base-pairing rule 1947 Erwin Chargaff, analysis of DNA from different species %A = %T & %C = %G Human DNA A = 30.9% T = 29.4% C = 19.9% G = 19.8%
14 Base-pairing in DNA
15 DNA Structure & Function 1. Historical perspective (DNA as the genetic material): Genetic transformation and DNA DNA is the genetic material in bacterial viruses (phage) The base-pairing rule DNA structure 2. Basis for polarity of SS DNA, and anti-parallel complementary strands of DNA 3. DNA replication models 4. Mechanism of DNA replication: steps and molecular machinery 5. Replication and the end of linear chromosomes- Molecular basis for aging 6. Fidelity of DNA replication
16 Structural Model of DNA Maurice Wilkins and Rosalind Franklin- X-ray crystallography (a) Rosalind Franklin (b) Franklin s X-ray diffraction Photograph of DNA
17 Watson and Crick deduced that DNA was a double helix Through observations of the X-ray crystallographic images of DNA
18 Watson and Crick - Specificity of pairing is dictated by the structure of the bases
19 Three models for DNA replication: Conservative model Semi-conservative model Dispersive model
20 Meselson-Stahl experiment Bacteria cultured in medium containing 15 N Bacteria transferred to medium containing 14 N DNA sample centrifuged after 20 min (after first replication) DNA sample centrifuged after 40 min (after second replication) Less dense More dense Check animation posted on course website
21 The Basic concept of DNA replication Each strand of DNA act as a template for synthesis of new complementary strands
22 Major Events in the History of Earth Cenozoic Humans Land plants Animals Origin of solar system and Earth RNA Multicellular eukaryotes 1 Proterozoiceon Archaeaneon Single-celled eukaryotes Atmospheric oxygen Prokaryotes
23 Molecular Mechanism of DNA Replication Collective action of several macro-molecules: DNA Proteins (enzymes & others) RNA Ribo-protein (for linear chromosomes) Direction of replication of new strands; How nucleotides are added in DNA replication Campbell Bio Flix: DNA Replication
24 DNA polymerase adds deoxyribonucleotides in a 5 to 3 direction, it adds nucleotides to the 3 end of a growing strand. The new strand always starts with the 5 end, the template starts with the 3 end.
25 Primase, an RNA polymerase, uses the DNA template strand to polymerize a short complementary RNA chain (RNA primer) Two different DNA polymerases both - cannot initiate the synthesis of a polynucleotide - can only add nucleotides to an existing 3 end
26 Summary of DNA Replication Semi-conservative Initiation: Origin of replication Primase and RNA primer Template strand vs. new strand 5 to 3 direction DNA polymerase (III and I) Base-pairing rules dntps: deoxy-atp, deoxy-gtp, deoxy-ctp, deoxy- TTP Leading and lagging strands Okazaki fragments DNA ligase Bidirectional Fidelity of DNA replication is maintained by activity of DNA polymerase and other proof reading systems.
27 Origin of Replication
28 Other proteins participate in DNA replication including: Helicase, topoisomerase, singlestrand binding protein
29 Replication of long DNA molecules begins at multiple origins of replication simultaneously and is bidirectional
30 Replicating the Ends of linear DNA Molecules Mechanism of DNA replication causes telomeres to get shorter with each round of replication org/courses/biolog y/units/cancer/ima ges.html RNA primer Lagging strand 5 3 Primer removed but cannot be replaced with DNA because no 3 end available for DNA polymerase 3 Last fragment 5 5 New leading strand 3 New leading strand 5 3 Previous fragment Removal of primers and replacement with DNA where a 3 end is available Second round of replication Further rounds of replication Shorter and shorter daughter molecules
31 Current Connections to DNA structure and replication Q: Why are we mortal with a limited life span? A: Our cells have a limited life span (# of cell divisions)
32 Telomerase- an enzyme (riboprotein) that extends the 3 end of the DNA strand by adding a repeated sequence of 6-nucleotides typically TTAGGG ( times) donald.slish/telomerase.html
33 Ends of linear chromosomes have special DNA sequences and are known as telomeres added by an enzyme known as telomerase after DNA replication is completed
34 Life span of dividing cells Telomerase is active in sperm, eggs, stem cells (bone marrow), and cancer cells but not in somatic tissues Most cells lose endmost bases after each cell division After about 50 divisions, shortened telomeres signal the cell to stop dividing Figure 2.3
35 Fidelity of DNA replication & maintaining DNA integrity Maintained by: 1. Proof-reading function of DNA polymerase 2. DNA repair systems DNA damage and repair in general
36 Genetic Integrity and Diversity Need for maintaining genetic integrity is balanced by having enough genetic variability for natural selection to act on Few errors of DNA replication are not corrected!
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