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DNA
             The Genetic Material




AP Biology                          2006-2007
Scientific History
     The march to understanding that DNA is
        the genetic material
          T.H. Morgan (1908)
          Frederick Griffith (1928)

          Avery, McCarty & MacLeod (1944)

          Erwin Chargaff (1947)

          Hershey & Chase (1952)

          Watson & Crick (1953)

          Meselson & Stahl (1958)


AP Biology
1908 | 1933
   Chromosomes related to phenotype
    T.H. Morgan
            working with Drosophila
              fruit flies
            associated phenotype with
             specific chromosome
              white-eyed male had specific
               X chromosome




AP Biology
1908 | 1933
   Genes are on chromosomes
    Morgan’s conclusions
         genes are on chromosomes
         but is it the protein or the

          DNA of the chromosomes
          that are the genes?
              initially proteins were thought
              to be genetic material…
              Why?

                 What’s so impressive
                  about proteins?!


AP Biology
1928
   The “Transforming Principle”
    Frederick Griffith
            Streptococcus pneumonia bacteria
              was working to find cure for pneumonia
         harmless live bacteria (“rough”)
          mixed with heat-killed pathogenic
          bacteria (“smooth”) causes fatal
          disease in mice
         a substance passed from dead

          bacteria to live bacteria to change
          their phenotype
                “Transforming Principle”

AP Biology
The “Transforming Principle” mix heat-killed
                                                                pathogenic &
live pathogenic      live non-pathogenic heat-killed            non-pathogenic
strain of bacteria   strain of bacteria   pathogenic bacteria   bacteria
A.                   B.                  C.                           D.




mice die              mice live           mice live         mice die




   Transformation = change in phenotype
   something in heat-killed bacteria could still transmit
   disease-causing properties
  AP Biology
1944
   DNA is the “Transforming Principle”
    Avery, McCarty & MacLeod
            purified both DNA & proteins separately from
             Streptococcus pneumonia bacteria
              which will transform non-pathogenic bacteria?
            injected protein into bacteria
              no effect
            injected DNA into bacteria
              transformed harmless bacteria into
               virulent bacteria

                                                      mice die

                   What’s the
AP Biology         conclusion?
1944 | ??!!
   Avery, McCarty & MacLeod
     Conclusion
            first experimental evidence that DNA was the
             genetic material




    Oswald Avery
AP Biology               Maclyn McCarty       Colin MacLeod
1952 | 1969
   Confirmation of DNA                              Hershey

     Hershey & Chase
          classic “blender” experiment
          worked with bacteriophage

               viruses that infect bacteria
             grew phage viruses in 2 media,
 Why use      radioactively labeled with either
  Sulfur       35S in their proteins
    vs.
Phosphorus?    32P in their DNA
             infected bacteria with
              labeled phages
AP Biology
Protein coat labeled                 DNA labeled with 32P
                         with 35S
Hershey                                    T2 bacteriophages
                                            are labeled with
& Chase                                   radioactive isotopes
                                                 S vs. P

                                          bacteriophages infect
                                             bacterial cells




                                    bacterial cells are agitated
Which                              to remove viral protein coats
radioactive
marker is found
inside the cell?

Which molecule
carries viral            35
                           S radioactivity            32
                                                        P radioactivity found
genetic info?
 AP Biology              found in the medium          in the bacterial cells
AP Biology
Blender experiment
     Radioactive phage & bacteria in blender
          35S phage
                 radioactive proteins stayed in supernatant
                 therefore viral protein did NOT enter bacteria
          32P phage
                 radioactive DNA stayed in pellet
                 therefore viral DNA did enter bacteria
            Confirmed DNA is “transforming factor”


                     Taaa-Daaa!
AP Biology
1952 | 1969
   Hershey & Chase                           Hershey




AP Biology   Martha Chase   Alfred Hershey
1947
   Chargaff
     DNA composition: “Chargaff’s rules”
          varies from species to species
          all 4 bases not in equal quantity

          bases present in characteristic ratio

              humans:
                         A = 30.9%      Rules
                                        A = T
                         T = 29.4%      C = G
                         G = 19.9%
                         C = 19.8%

                  That’s interesting!
                 What do you notice?
AP Biology
1953 | 1962
   Structure of DNA
     Watson & Crick
            developed double helix model of DNA
              other leading scientists working on question:
                Rosalind Franklin
                Maurice Wilkins
                Linus Pauling




AP Biology
              Franklin           Wilkins            Pauling
1953 article in Nature
  Watson and Crick




    Watson
AP Biology           Crick
Rosalind Franklin (1920-1958)




AP Biology
But how is DNA copied?
      Replication of DNA
            base pairing suggests
             that it will allow each
             side to serve as a
             template for a new
             strand




“It has not escaped our notice that the specific pairing we have postulated
immediately suggests a possible copying mechanism for the genetic
  AP Biology
material.”                                             — Watson & Crick
Can you design
                                               a nifty experiment
                                                    to verify?
    Models of DNA Replication
     Alternative models
            become experimental predictions

    conservative        semiconservative       dispersive

P


1


2
AP Biology
1958
   Semiconservative replication
     Meselson & Stahl
             label “parent” nucleotides in DNA strands with
              heavy nitrogen = 15N
             label new nucleotides with lighter isotope = 14N
                              “The Most Beautiful Experiment in Biology”

                              parent   replication
Make predictions…

                 15
                      N/15N



    15
         N parent
         strands
AP Biology
Predictions
                              14
                                   N/14N
 1st round of
                                           15
                                                N/14N    N/14N
                                                        15
                              15
                                   N/15N
 replication

                                      semi-
                      conservative              dispersive
                                   conservative
 2nd round of
 replication

P
                              14
                                   N/14N   14
                                             N/14N
                                           15
                                             N/14N      15
                                                             N/14N
                              15
                                   N/15N
1        15
              N/15N
                                      semi-
2 N parent
    15                conservative              dispersive
 AP Biology                        conservative
  strands
Meselson & Stahl
Matthew Meselson   Franklin Stahl



                                                       Franklin Stahl




                                    Matthew Meselson




AP Biology
Scientific History
      March to understanding that DNA is the genetic material
            T.H. Morgan (1908)
               genes are on chromosomes
            Frederick Griffith (1928)
               a transforming factor can change phenotype
            Avery, McCarty & MacLeod (1944)
               transforming factor is DNA
            Erwin Chargaff (1947)
               Chargaff rules: A = T, C = G
            Hershey & Chase (1952)
               confirmation that DNA is genetic material
            Watson & Crick (1953)
               determined double helix structure of DNA
            Meselson & Stahl (1958)
AP Biology     semi-conservative replication
The “Central Dogma”
     Flow of genetic information in a cell

              transcription         translation

   DNA                        RNA                 protein


         replication

AP Biology
Science …. Fun
               Party Time!
             Any Questions??




AP Biology                    2006-2007
Ghosts of Lectures Past
                    (storage)




AP Biology                             2006-2007
1958
   Semiconservative replication
     Meselson & Stahl
             label “parent” nucleotides in DNA strands with
              heavy nitrogen = 15N
             label new nucleotides with lighter isotope = 14N
                              “The Most Beautiful Experiment in Biology”

                              parent   replication


                 15
                      N/15N



    15
         N parent
         strands
AP Biology
1958
   Semiconservative replication
     Make predictions…
         15  N strands replicated in 14N medium
            1st round of replication? where should the bands be?
            2nd round?




AP Biology

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60 ch14dn ahistory2008

  • 1. DNA The Genetic Material AP Biology 2006-2007
  • 2. Scientific History  The march to understanding that DNA is the genetic material  T.H. Morgan (1908)  Frederick Griffith (1928)  Avery, McCarty & MacLeod (1944)  Erwin Chargaff (1947)  Hershey & Chase (1952)  Watson & Crick (1953)  Meselson & Stahl (1958) AP Biology
  • 3. 1908 | 1933 Chromosomes related to phenotype  T.H. Morgan  working with Drosophila  fruit flies  associated phenotype with specific chromosome  white-eyed male had specific X chromosome AP Biology
  • 4. 1908 | 1933 Genes are on chromosomes  Morgan’s conclusions  genes are on chromosomes  but is it the protein or the DNA of the chromosomes that are the genes?  initially proteins were thought to be genetic material… Why? What’s so impressive about proteins?! AP Biology
  • 5. 1928 The “Transforming Principle”  Frederick Griffith  Streptococcus pneumonia bacteria  was working to find cure for pneumonia  harmless live bacteria (“rough”) mixed with heat-killed pathogenic bacteria (“smooth”) causes fatal disease in mice  a substance passed from dead bacteria to live bacteria to change their phenotype  “Transforming Principle” AP Biology
  • 6. The “Transforming Principle” mix heat-killed pathogenic & live pathogenic live non-pathogenic heat-killed non-pathogenic strain of bacteria strain of bacteria pathogenic bacteria bacteria A. B. C. D. mice die mice live mice live mice die Transformation = change in phenotype something in heat-killed bacteria could still transmit disease-causing properties AP Biology
  • 7. 1944 DNA is the “Transforming Principle”  Avery, McCarty & MacLeod  purified both DNA & proteins separately from Streptococcus pneumonia bacteria  which will transform non-pathogenic bacteria?  injected protein into bacteria  no effect  injected DNA into bacteria  transformed harmless bacteria into virulent bacteria mice die What’s the AP Biology conclusion?
  • 8. 1944 | ??!! Avery, McCarty & MacLeod  Conclusion  first experimental evidence that DNA was the genetic material Oswald Avery AP Biology Maclyn McCarty Colin MacLeod
  • 9. 1952 | 1969 Confirmation of DNA Hershey  Hershey & Chase  classic “blender” experiment  worked with bacteriophage  viruses that infect bacteria  grew phage viruses in 2 media, Why use radioactively labeled with either Sulfur  35S in their proteins vs. Phosphorus?  32P in their DNA  infected bacteria with labeled phages AP Biology
  • 10. Protein coat labeled DNA labeled with 32P with 35S Hershey T2 bacteriophages are labeled with & Chase radioactive isotopes S vs. P bacteriophages infect bacterial cells bacterial cells are agitated Which to remove viral protein coats radioactive marker is found inside the cell? Which molecule carries viral 35 S radioactivity 32 P radioactivity found genetic info? AP Biology found in the medium in the bacterial cells
  • 12. Blender experiment  Radioactive phage & bacteria in blender  35S phage  radioactive proteins stayed in supernatant  therefore viral protein did NOT enter bacteria  32P phage  radioactive DNA stayed in pellet  therefore viral DNA did enter bacteria  Confirmed DNA is “transforming factor” Taaa-Daaa! AP Biology
  • 13. 1952 | 1969 Hershey & Chase Hershey AP Biology Martha Chase Alfred Hershey
  • 14. 1947 Chargaff  DNA composition: “Chargaff’s rules”  varies from species to species  all 4 bases not in equal quantity  bases present in characteristic ratio  humans: A = 30.9% Rules A = T T = 29.4% C = G G = 19.9% C = 19.8% That’s interesting! What do you notice? AP Biology
  • 15. 1953 | 1962 Structure of DNA  Watson & Crick  developed double helix model of DNA  other leading scientists working on question:  Rosalind Franklin  Maurice Wilkins  Linus Pauling AP Biology Franklin Wilkins Pauling
  • 16. 1953 article in Nature Watson and Crick Watson AP Biology Crick
  • 18. But how is DNA copied?  Replication of DNA  base pairing suggests that it will allow each side to serve as a template for a new strand “It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic AP Biology material.” — Watson & Crick
  • 19. Can you design a nifty experiment to verify? Models of DNA Replication  Alternative models  become experimental predictions conservative semiconservative dispersive P 1 2 AP Biology
  • 20. 1958 Semiconservative replication  Meselson & Stahl  label “parent” nucleotides in DNA strands with heavy nitrogen = 15N  label new nucleotides with lighter isotope = 14N “The Most Beautiful Experiment in Biology” parent replication Make predictions… 15 N/15N 15 N parent strands AP Biology
  • 21. Predictions 14 N/14N 1st round of 15 N/14N N/14N 15 15 N/15N replication semi- conservative dispersive conservative 2nd round of replication P 14 N/14N 14 N/14N 15 N/14N 15 N/14N 15 N/15N 1 15 N/15N semi- 2 N parent 15 conservative dispersive AP Biology conservative strands
  • 22. Meselson & Stahl Matthew Meselson Franklin Stahl Franklin Stahl Matthew Meselson AP Biology
  • 23. Scientific History  March to understanding that DNA is the genetic material  T.H. Morgan (1908)  genes are on chromosomes  Frederick Griffith (1928)  a transforming factor can change phenotype  Avery, McCarty & MacLeod (1944)  transforming factor is DNA  Erwin Chargaff (1947)  Chargaff rules: A = T, C = G  Hershey & Chase (1952)  confirmation that DNA is genetic material  Watson & Crick (1953)  determined double helix structure of DNA  Meselson & Stahl (1958) AP Biology  semi-conservative replication
  • 24. The “Central Dogma”  Flow of genetic information in a cell transcription translation DNA RNA protein replication AP Biology
  • 25. Science …. Fun Party Time! Any Questions?? AP Biology 2006-2007
  • 26. Ghosts of Lectures Past (storage) AP Biology 2006-2007
  • 27. 1958 Semiconservative replication  Meselson & Stahl  label “parent” nucleotides in DNA strands with heavy nitrogen = 15N  label new nucleotides with lighter isotope = 14N “The Most Beautiful Experiment in Biology” parent replication 15 N/15N 15 N parent strands AP Biology
  • 28. 1958 Semiconservative replication  Make predictions…  15 N strands replicated in 14N medium  1st round of replication? where should the bands be?  2nd round? AP Biology

Editor's Notes

  • #6: Fred Griffith, English microbiologist, dies in the Blitz in London in 1941
  • #8: 1. Purified S strain extracts to characterize the transforming principle. 2. Material was resistant to proteases; it contained no lipid or carbohydrate. 3. If DNA in the extract is destroyed, the transforming principle is lost. 4. Pure DNA isolated from the S strain extract transforms R strain. 5. Avery cautiously suggested that DNA was the genetic material. 6. This was the first experimental evidence that DNA is the genetic material.
  • #9: Maclyn McCarty (June 9, 1911 – January 2, 2005) was an American geneticist. Oswald Avery (October 21, 1877–2 February 1955) was a Canadian-born American physician and medical researcher. Colin Munro MacLeod (January 28, 1909 — February 11, 1972) was a Canadian-American geneticist. After Oswald T. Avery, Colin M. MacLeod, and Maclyn McCarty published the 1944 article, a number of their contemporaries immediately understood that transformation was the transfer of genetic material from one bacterium to another, and that the transforming substance, DNA, must be the genetic material. However, the team's somewhat tentatively stated conclusions were not met with complete acceptance. At the time, the belief that DNA was a monotonous chain of four repeating nucleotides--structurally important but of little physiological interest--was still difficult to overcome. The belief that only proteins possessed the structural complexity necessary to carry hereditary information was pervasive among geneticists. Many of the scientists who had previously thought that genetic material was protein still believed that the effects of the transforming principle were perhaps due to some undetected protein associated with the DNA.
  • #14: Martha Cowles Chase (1927 – August 8, 2003) was a young laboratory assistant in the early 1950s when she and Alfred Hershey conducted one of the most famous experiments in 20th century biology. Devised by American bacteriophage expert Alfred Hershey at Cold Spring Harbor Laboratory New York, the famous experiment demonstrated the genetic properties of DNA over proteins. By marking bacteriophages with radioactive isotopes, Hershey and Chase were able to trace protein and DNA to determine which is the molecule of heredity. Hershey and Chase announced their results in a 1952 paper. The experiment inspired American researcher James D. Watson, who along with England's Francis Crick figured out the structure of DNA at the Cavendish Laboratory of the University of Cambridge the following year. Hershey shared the 1969 Nobel Prize in Physiology or Medicine with Salvador Luria and Max Delbrück. Chase, however, did not reap such rewards for her role. A graduate of The College of Wooster in Ohio (she had grown up in Shaker Heights, Ohio), she continued working as a laboratory assistant, first at the Oak Ridge National Laboratory in Tennessee and then at the University of Rochester before moving to Los Angeles in the late 1950s. There she married biologist Richard Epstein and earned her Ph.D. in 1964 from the University of Southern California. A series of personal setbacks through the 1960s ended her career in science. She spent decades suffering from a form of dementia that robbed her of short-term memory. She died on August 8, 2003.
  • #16: Watson & Crick’s model was inspired by 3 recent discoveries: Chargaff’s rules Pauling’s alpha helical structure of a protein X-ray crystallography data from Franklin & Wilkins
  • #18: A chemist by training, Franklin had made original and essential contributions to the understanding of the structure of graphite and other carbon compounds even before her appointment to King's College. Unfortunately, her reputation did not precede her. James Watson's unflattering portrayal of Franklin in his account of the discovery of DNA's structure, entitled "The Double Helix," depicts Franklin as an underling of Maurice Wilkins, when in fact Wilkins and Franklin were peers in the Randall laboratory. And it was Franklin alone whom Randall had given the task of elucidating DNA's structure. The technique with which Rosalind Franklin set out to do this is called X-ray crystallography. With this technique, the locations of atoms in any crystal can be precisely mapped by looking at the image of the crystal under an X-ray beam. By the early 1950s, scientists were just learning how to use this technique to study biological molecules. Rosalind Franklin applied her chemist's expertise to the unwieldy DNA molecule. After complicated analysis, she discovered (and was the first to state) that the sugar-phosphate backbone of DNA lies on the outside of the molecule. She also elucidated the basic helical structure of the molecule. After Randall presented Franklin's data and her unpublished conclusions at a routine seminar, her work was provided - without Randall's knowledge - to her competitors at Cambridge University, Watson and Crick. The scientists used her data and that of other scientists to build their ultimately correct and detailed description of DNA's structure in 1953. Franklin was not bitter, but pleased, and set out to publish a corroborating report of the Watson-Crick model. Her career was eventually cut short by illness. It is a tremendous shame that Franklin did not receive due credit for her essential role in this discovery, either during her lifetime or after her untimely death at age 37 due to cancer.
  • #29: Matthew Stanley Meselson (b. May 24, 1930) is an American geneticist and molecular biologist whose research was important in showing how DNA replicates, recombines and is repaired in cells. In his mature years, he has been an active chemical and biological weapons activist and consultant. He is married to the medical anthropologist and biological weapons writer Jeanne Guillemin. Dr. Franklin William Stahl (born October 8, 1929) is an American molecular biologist. With Matthew Meselson, Stahl conducted the famous Meselson-Stahl experiment showing that DNA is replicated by a semiconservative mechanism, meaning that each strand of the DNA serves as a template for the "replicated" strand. He is Emeritus Professor of Biology[1] at the University of Oregon's Institute of Molecular Biology in Eugene, Oregon.