Life begins with a cell

• We are made of trillions cells

• We can grow, reproduce, process information,
respond to stimuli and our body carry an
amazing array of chemical activity
• Many unicellular organisms also have all
hallmark properties of life
• Indicating the cell as unit of life
• Birth, life and death of cells
• Life begins with a cell
 Cells and diversity
• Amazing variety of sizes and shapes, mobile
and sessile, oxygen kills some others are killed
without oxygen. But all are cells.
• Cells are prokaryotic or eukaryotic (presumed to
have common progenitor with archaea)
• Prokaryotic cells include bacteria and archaea.
They include a simple compartment surrounded
by pm
• They lack a defined Nucleus
Changes in cells underlie evolution
• Prokaryotes are small microscopic organisms
• Single E. coli has dry wt of 25 x 10 -14g yet
bacteria account for 1-1.5 Kg average human wt.
• Estimated number of bacteria on earth is 5x1030,
weighing about 1012 kg. Fast reproduction
• Some species have changed little over a time
and others have changed dramatically
• These changes affected by environment that
causes increased survival/demise of variants
• Both stasis and change are possible :. of
precise copying of genetic material, yet
its rare errors introduce variations
 Primary Genetic Material - for
Evolution
• Carries within its st. hereditary information that
determines the st. of proteins, it is the prime substance.
• Instructions are encoded that direct cells to grow and
divide via messages that makes zygote to differentiate to
multitude of specialized cells
• Genetic material allows infinite variations and is basis of
evolution resulting in different life forms in 3-4 billion
years ago since first life forms appeared on earth
• Biology has come of age in last fifty years; informed
minds say life can be explained by laws physics and
mathematics – like why apple falls and moon does not
Mendel’s breeding experiments 1856
• Selective breeding – crop, animals - empirical
• Selective pea plant breeding scientifically
• Pure breeding - contrasting traits
• No blending of traits - independent assortment
• Dominant and recessive
• F1 progeny of pure bred gave dominant offspring
• Interbreeding F1 progeny reveled 3:1 in F2
• Cross bred plants with two contrasting features in
F2 gave 9:3:3:1
Mendel’s seminal work published 1865
to be rediscovered in 1900
• Ratio could be accounted for by assuming that each
plant posses two factors for each character
• During gamete formation (sex) two traits separate so that
each gamete receive one trait
• Gamete: equal chance of dominant or recessive
• William Bateson (UK) gave the name genetics 1906 and
Wilhelm Johannsen coined gene 1909
• Each gene can exist in different forms or alleles
• Both alleles of gene are same individual is homozygous
• Genotype is complete genetic composition of an
organism, whereas phenotype is the physical expression
of the genotype.
 Chromosomes: bearers of heredity
• Advances made in microscopy in late 19th century
enabled to study cell structure
• Nucleus attracted attention as it disappeared before cell
division and chromosomes appeared
• Walter Sutton 1902 described chromosomes of
grasshopper and drew striking parallel b/w these and
Mendel’s factors
• Ch # was doubled during cell division (mitosis) 2n,
producing two daughter cells
• Ch# was shown to be half in sex cells (meiosis) 1n
• Fertilizations restores 2n Ch #
 Genes mapped to chromosomes
• First trait assigned to chromosome (ch) was sex
• 1905 in Columbia Univ observed blowfly ch. X ch present in two
copies in females & only one copy in male somatic cells which also
carries a morphologically distinct Y ch. Dad made us a boy or a
girl
• Second in 1910 also from Columbia Univ by TH Morgan eye color of
fruit fly
• Linkage groups, Four ch. In Drosophila, linkage maps, distance
b/w pair of genes over which crossing-over occurs once in 100
meiosis was named as centimorgan
• Recombination frequencies measure genetic distances and may
differ from physical distances b/w genes. This is particularly true for
genes separated by large physical distances
 Mutations
• Spontaneous mutants result from changes in single
genes and genetically stable
• Like white eyed Drosophila Vs wild type. Genetic flags
• They occur at low frequency. Relief came
• 1926 Herman Muller found 100 X-rayed-induced
Drosophila mutants in 2 months, compared 200 mutants
found over previous 16 yrs
• Finding had profound implications on human health –
dangers of unshielded ionizing radiation source
• Likewise many chemicals are identified as mutagens
• Mustard gas, ethyl methanesulfonate, ethylnitrosourea
• ENS Has been used to induce mutations in Zebra Fish
• Genetic screens through mutations
Chromosomes contain protein & nucleic acid
• Frederick Griffith 1928(UK) working with Diplococcus
pneumoniae made an observation that heat killed
pathogenic cells mixed with nonpathogenic cells and
injected to mice were able to transform a small % of
nonpathogenic cells to pathogenic cells. In becoming
pathogenic the non-virulent cells acquired thick, outer,
polysaccharide-rich cell wall. Griffith did not identify the
substance
• Oswald Avery at RU worked on ‘transforming factor’
• It took 10 yrs to prove DNA is transforming factor – 1944 -
which is destroyed using a then just discovered DNase
whereas transforming activity remained unaffected using
RNase or proteases.
• Skeptics - “genetic proteins”
Avery-MacLeod-McCarty Experiment
 Viruses and phages
• In 1930 it became possible to purify viruses
including bacteriophages
• Nature of these tiny particles remain disputed
• EM showed they were not minute cells
• Appeared as protein packages containing NA
• Best guess: obligatory parasites at genetic level
• Phages multiply within E. coli and need 20
minutes to produce several hundred progeny
• Good material for genetic/mutant studies
Hershey Chase Experiment 1952: Phage DNA
alone carries genetic information
 Gene concept to reality
• William Bateson (UK) gave the name genetics 1906 and Wilhelm
Johannsen coined gene 1909
• 1942 G. Beadle & E. Tatum found X-ray induced mutations in fungi
often caused specific biochemical defect, reflecting absence or
malfunctioning of enzyme
• This led to one gene one enzyme hypothesis.
• This has to be expanded to accommodate discoveries like not all
genes encode enzymes, many encode proteins, some do not
encode protein, some encode multiple proteins (alternate splicing).
Both in pro and eukaryotes multiple gene products can be
generated by alternate promoter or polyadenylation site usage.
• In more obscure cases two or more genes may be required to
generate a single polypeptide – trans-splicing
 What genes made up of

• Chemical nature of chromosomes long predated any knowledge of

their role in heredity

• 1869 Friedrich Miescher, at Tubingen isolated P containing material

called nuclein from degenerate WBC in pus (plentiful in days prior to
antibiotics)
• 1874 he found salmon sperm better for isolating nuclein. He
showed it be a mix of +ly charged proteins & acid material that was
named in1889 by Richard Altman as nucleic acid

• By degrading NA German chemist Albrecht Kossel discovered N

containing flat “bases” guanine (1882), adenine (1886), thymine
(1893), cytosine (1894) and uracil (1900)
• Phoebus Levene at RU identified deoxyribose in DNA and ribose in
RNA
 Quantification of DNA bases
• In 1940s it became possible to quantify nitrogen bases using
chromatographic methods developed for aa in a protein by Martin and
Synge
• Bases of DNA were first quantitatively analyzed in lab of Erwin Chargaff
at Columbia Uni
• By 1951 it was clear that 4 bases were not present in equal amount. It also
meant the potential # of different DNA sequences was astronomically
large 4n where n is the # of nucleotides in a DNA
• Chargaff failed to realize that A/T and G/C ratios held clues to DNA st
• Quantitative determination of DNA amounts in cells supported it to be
genetic material as amount was same in somatic cells, half in gamete cells
• Equally important was A+T and G+C was not similar for all organisms
• He analyzed DNA from human sperms, ox thymus and yeast
 Alexander Todd Cambridge Univ
• EM studies on DNA: extremely
elongated molecules thousands of
Ao in length and approximately 20
Ao thick
• Alexander Todd in 1952 showed
that phosphate-ester linkages
bound nucleotides together. P
connecting the 5/ carbon atom of
one deoxyribose residue to the 3/
C of deoxyribose in the adjacent
nucleotide
• This means DNA strand ends with
5/ P gp at one end and 3/ OH gp at
the other. Todd concluded DNA is
linear molecule
• Thee years later RNA was found
to have regular back bone and has
5/- 3/ phosphodiester links
 Rosalind Franklin
• DNA exposed to high RH (paracrystalline, pattern “B”
form) was seen in 1951 by English physical chemist
Rosalind Franklin a colleague of Wilkins in Kings College
• Her pictures revealed a dominant cross-link pattern, the
telltale mark of a helix
• A separate nucleotide was found every 3.4 Ao along its
fiber axis, with 10 nucleotides, or 34 Ao, being required
for every turn of the helix
• 20 Ao breadth was too large to contain one chain
• X ray diffraction favored two chains that ran in opposite
directions
 Model of DNA – Watson & Crick
• James Watson & Francis Crick, Cavendish Lab of Cambridge Uni
1953 built 3D model of DNA in which P-Sug backbones are outside,
purine and pyrimidine are inside, oriented so that bases on one
strand can form hydrogen bonds to those bases on opposite chains

• Complementary polynucleotide chains are held by several hydrogen

bonds thus holding them firmly. A=T, G =C
• Purine (A or G) of one chain pairs with Pyrimidine (T or C) of other
• Earlier anomaly that T2 phage does not have cytosine provided
support as methyl cytosine was instead discovered in the phage
• Sequence of one chain is known than its partner is also known
• Symmetry of two strands is antiparallel
• Promptly proposed that two strands be regarded as + and – of helix
each template specifying its complement and thereby capable of
generating two daughter DNA molecules
 DNA replication: semi-conservative
• What was not known the process whether conservative, in which
original ds stays together and new complementary strands form a
new helix, or semi-conservative, where the two original strands
separate completely during the synthesis of new complementary
strands, so new double helix contains one parental and one new
strand
• It was in 1958 Meselson and Stahl cleverly used density difference
to separate parental DNA mol from daughter mol to show that DNA
replication is semiconservative. ( E. coli grown in 15NH4Cl & 14NH4Cl)
• Independently Arthur Kornberg’s lab working on cell free extracts
inferred that DNA chain serve as direct templates for the formation
of new DNA chains.
• DNA III polymerase was isolated by Tom Kornberg
• Denaturing and renaturing of DNA, hybridization
• Chromosomes are very long DNA molecules
• Some viruses have an RNA genome
 Historical overview
1. Life begins with a cell
2. Diversity and commonality of cells
3. Changes in cells underlie evolution
4. Primary Genetc Material - for Evolution
5. Mendel’s breeding experiments 1856
6. Mendel’s seminal work publishied 1865
7. Chromosomes: bearers of heredity
8. Genes mapped to chromosomes
9. Mutations
10. Chromosomes contain protein & nucleic acid
11. Avery-MacLeod-McCarty Experiment
12. Viruses and phages
13. Hershey Chase Experiment 1952
14. Gene concept to reality
15. What genes made up of
16. Quantification of DNA bases
17. Alexander Todd Cambridge Univ
18. Rosalind Franklin
19. Model of DNA – Watson & Crick
20. DNA replication: semi-conservative
• How four letter linear language of DNA is translated into 20-letter
language of polypeptide chains?