- •In this chapter we will:
- •Structure of dna 195
- •Discovering the Structure of dna
- •Information, is a double helical molecule.
- •Polymerase Chain Reaction (pcr)
- •Replication of dna 201
- •Ames Test
- •One Gene—One Polypeptide
- •Expression of genetic information 207
- •212 Chapter 7 microbial genetics: replication and expression of genetic information
FIG.
7-4 Four different deoxyribonucleotides comprise the subunit
molecules of DNA. These have differing nucleic acid bases:
thymine, cytosine, adenine, and guanine.
FIG.
7-5 Nucleotides are joined together by phos- phodiester bonds
between the 3'-OH and 5'-P positions. There is a free 5'-P at
one end of the polynucleotide chain and a free 3'- OH at the
other end.
Structure of dna 195
196 CHAPTER 7 MICROBIAL GENETICS: REPLICATION AND EXPRESSION OF GENETIC INFORMATION
phate
forms a bridge between the number 3-carbon of one deoxyribose sugar
molecule and the number 5-carbon of another. At one end of the chain
there is no phosphate bonded to the 3-carbon of a deoxyribose,
and at the other end of the molecule, the phosphate attached to
the 5-carbon is not involved in forming a phosphodiester linkage.
Thus there is a free hydroxyl group at the 3-carbon position at one
end of the chain (3'-OH
free end)
and
there is a free phosphate group at the 5-carbon position at the
other end (5'-P
free end).
This
imparts a second important property on the DNA macromolecule—that
of directionality.
Having
different groups free at the different ends of the molecule
distinguishes one end from the other (like left from right) thereby
permitting the molecule to be read from a particular direction. This
is especially critical for a molecule whose purpose is to store
and to transmit genetic information because it estab
lishes
the basis for the correct direction for reading the order of
nucleotides that encode the genetic information within the DNA
macromolecule.
The
chains of DNA macromolecules are different at
either
end, which allows directional recognition.
DNA
Double
Helix—Complementarity
There
are two chains of nucleotides in the DNA macromolecule. The two
polynucleotide chains that comprise the DNA double helix run in
opposite directions—one chain runs from the 3'-OH to the 5'-P
free end and the complementary chain runs from the 5'-P to the 3'-OH
free end. These complementary chains twist together to form an
arrangement called a double
helix
(FIG.
7-6).
The
two chains of the DNA double helix are held together by hydrogen
bonding between complementary chains. Two of the nitrogenous
bases (C and T)
FIG.
7-6 The
double helix is the fundamental structure of the DNA macromolecule.
The two strands are held together by hydrogen bonding between
complementary base pairs. There are three hydrogen bonds between the
base pairs guanine and cytosine and two hydrogen bonds between
the base pairs adenine and thymine.
In
the early 1950s, James Watson, who had only recently received a
doctoral degree from Indiana University, teamed with Francis Crick,
a Cambridge University researcher, and pieced together the
available data to determine the chemical structure of the DNA
molecule, showing how DNA could store and transmit hereditary
information (see Figure).
Watson
and Crick reasoned that the double-ringed adenine and guanine
molecules were probably paired with the single-ringed thymine and
cytosine molecules along the entire length of DNA. This would fit
the observation made by Erwin Chargaff in the United States in
the late 1940s that in any given DNA macromolecule the amount of
adenine present is always equal to the amount of thymine and the
amount of cytosine is always equal to the amount of guanine. It
also was consistent with the observations of Maurice Wilkins
and Rosalind Franklin at Cambridge University in England, who, using
X-ray diffraction methods, showed that DNA was long and thin, with a
uniform diameter. If this was not the case, then DNA would bulge
where the two double rings were paired and narrow where two single
rings were paired, something that was never seen in the X-ray
diffraction images of Wilkins and Franklin.
The
problem Watson and Crick had was that the structural formulas for
the nucleic acid bases guanine and thymine that were in the organic
chemistry books at that time were wrong. Because they were trying to
build a model with the wrong structural representations for these
molecules, they were unable to get the molecules to fit in a
way that would be stabilized by hydrogen bonding between their
atoms. Only after getting the structural forms of guanine and
thymine corrected during a chance meeting with a visiting
scientist were Watson and Crick able to build the correct model
of DNA. Once they had the accurate structures, they began shifting
the bases in and out of various pairing possibilities.
They
next used a plumb line and a measuring stick to determine the
relative positions of all of the atoms in a single nucleotide. By
assuming a helical shape like a spiral staircase, it became clear
that the locations of the atoms in one nucleotide would
automatically generate the position of the other. They had
constructed a model of the DNA double helix. The model Watson and
Crick constructed also showed how the chains could be separated
and how each could code for a new complementary chain, thereby
unraveling the mystery of how a chemical could pass hereditary
information to the next generation with such fidelity.
James
Watson at age 23 and Francis Crick at age 34 developed a model for
the structure of DNA while working at the Cavendish Laboratory at
Cambridge University, England. The model explained how DNA can
transmit hereditary information. They announced their discovery of
the molecular structure of DNA in 1953 and shared the Nobel prize
for medicine in 1962 along with Maurice Wilkins.
In
1993, on the fortieth anniversary of their discovery, they again
posed with their model of DNA, which has proven to be correct.
Discovering the Structure of dna
