DNA

MODELO DEL ADN

SECUENCIACION DE GENES
Introduction to DNA Structure
MICROFOTOGRAFIA ELECTRONICA
FLUJO DE INFORMACION GENETICA
DNA
TIPOS DE CELULAS
NIVELES DE
ORGANIZACION
GENES
VIRUS
 DNA Double Helix

DNA is a normally double stranded macromolecule. Two

polynucleotide chains, held together by weak thermodynamic
forces, form a DNA molecule.
DNA Double Helix
DNA Double Helix
DNA
 Components of DNA

DNA is a polymer. The monomer units of DNA are nucleotides, and the
polymer is known as a "polynucleotide." Each nucleotide consists of a 5-
carbon sugar (deoxyribose), a nitrogen containing base attached to the
sugar, and a phosphate group. There are four different types of
nucleotides found in DNA, differing only in the nitrogenous base. The
four nucleotides are given one letter abbreviations as shorthand for the
four bases.

•A is for adenine
•G is for guanine
•C is for cytosine
•T is for thymine
ESTRUCTURA DNA
ESTRUCTURA DNA
Purine Bases
Adenine and guanine are purines. Purines are the larger of the
two types of bases found in DNA. Structures are shown below:
Structure of A and G

The 9 atoms that make up the fused rings (5 carbon, 4 nitrogen)

are numbered 1-9. All ring atoms lie in the same plane.
 Pyrimidine Bases
Cytosine and thymine are pyrimidines. The 6 stoms (4 carbon, 2 nitrogen)
are numbered 1-6. Like purines, all pyrimidine ring atoms lie in the same
plane.
Structure of C and T
Deoxyribose Sugar

The deoxyribose sugar of the DNA backbone has 5 carbons and 3

oxygens. The carbon atoms are numbered 1', 2', 3', 4', and 5' to
distinguish from the numbering of the atoms of the purine and
pyrmidine rings. The hydroxyl groups on the 5'- and 3'- carbons link to
the phosphate groups to form the DNA backbone. Deoxyribose lacks an
hydroxyl group at the 2'-position when compared to ribose, the sugar
component of RNA.
Structure of deoxyribose
Nucleosides

A nucleoside is one of the four DNA bases covalently attached to the C1'
position of a sugar. The sugar in deoxynucleosides is 2'-deoxyribose. The
sugar in ribonucleosides is ribose. Nucleosides differ from nucleotides in
that they lack phosphate groups. The four different nucleosides of DNA
are deoxyadenosine (dA), deoxyguanosine (dG), deoxycytosine (dC), and
(deoxy)thymidine (dT, or T).
 Structure of dA

In dA and dG, there is an "N-glycoside" bond between the sugar C1'

and N9 of the purine.
Nucleotides

A nucleotide is a nucleoside with one or more phosphate groups covalently

attached to the 3'- and/or 5'-hydroxyl group(s).

DNA Backbone

The DNA backbone is a polymer with an alternating sugar-phosphate

sequence. The deoxyribose sugars are joined at both the 3'-hydroxyl and 5'-
hydroxyl groups to phosphate groups in ester links, also known as
"phosphodiester" bonds
Example of DNA Backbone: 5'-d(CGAAT):
Features of the 5'-d(CGAAT) structure:

•Alternating backbone of deoxyribose and phosphodiester groups

•Chain has a direction (known as polarity), 5'- to 3'- from top to bottom

•Oxygens (red atoms) of phosphates are polar and negatively

charged

•A, G, C, and T bases can extend away from chain, and stack atop
each other

•Bases are hydrophobic

Structure of DNA Double Helix
 Features of the DNA Double Helix

•Two DNA strands form a helical spiral, winding around a helix

axis in a right-handed spiral

•The two polynucleotide chains run in opposite directions

•The sugar-phosphate backbones of the two DNA strands wind

around the helix axis like the railing of a sprial staircase

•The bases of the individual nucleotides are on the inside of the

helix, stacked on top of each other like the steps of a spiral
staircase.
Base Pairs
Within the DNA double helix, A forms 2 hydrogen bonds with T on the opposite
strand, and G forms 3 hyrdorgen bonds with C on the opposite strand.

Example of dA-dT base pair as found within DNA double

Example of dG-dC base pair as found within DNA double helix
•dA-dT and dG-dC base pairs are the same length, and
occupy the same space within a DNA double helix.
Therefore the DNA molecule has a uniform diameter.

•dA-dT and dG-dC base pairs can occur in any order within
DNA molecules
 DNA Helix Axis

The helix axis is most apparent from a view directly down the axis. The sugar-
phosphate backbone is on the outside of the helix where the polar phosphate
groups (red and yellow atoms) can interact with the polar environment. The
nitrogen (blue atoms) containing bases are inside, stacking perpendicular to the
helix axis.
View down the helix axis
NUCLEOIDE
 How Small Are Bacteria?

Magnified
14,000 times

Bacillus cells on the tip of a pin

 Microbial genomes: What are the sources of genetic variation?
1. Arber, W. (2000) Genetic variation: molecular mechanisms & impact on microbial evolution.
FEMS Microbiology Rev. 24: 1.
2. Levin, B. R. and C. T. Bergstrom (2000) Bacteria are different: Observations, interpretations,
speculations and opinions about mechanisms of adaptive evolution in prokaryotes. PNAS 97: 6981.

I. Reading a scientific paper.

A. Procedural issues
B. Conceptual issues
C. Being critical

II. Arber’s thesis: Prokaryotes have

“evolution genes.”

III. Strong or weak evidence?

DNA BACTERIANO
E. coli 0157
 Introduction
The Central Dogma
of Molecular Biology

Cell

Transcription DNA

mRNA
Translation Ribosome

Polypeptide
(protein)
ESTRUCTURA DNA
M UTAC I O N E S
 ¿ Qué son las mutaciones ?

Mutations and Mutagenesis

•A mutation is a change in the DNA sequence of a chromosome.

–note: a mutation involves a change in a nucleotide pair compared to wild

type.

•Can arise spontaneously without an identifiable cause.

•Can arise by mutagens-UV radiation, chemicals that modify bases
(nitrous acid -deaminatedcytosine and adenine).
•A mutant is a organism that has a mutation.
Cuando se produce una mutación, la nueva forma del gen se
hereda de una manera estable, justamente como la forma que le
precedió.
 El organismo que lleva el gen alterado se llama Mutante, al que
lleva el gen normal (inalterado) se le conoce como “tipo silvestre”.

 se dice que la mayoría de las mutaciones dañan la función de una

proteína, por tal motivo su estudio se extiende a analizar las
situaciones en que un gen no funciona adecuadamente.
 El efecto fenotípico de un cambio en una proteína depende de su
función en el organismo, puede variar desde un efecto nulo a un efecto
letal. Cambios complejos en el fenotipo pueden resultar de una sola
mutación .

 Una mutación en el gen puede alterar la actividad de la proteína de la

cual él es responsable, esto explica la naturaleza de las mutaciones
recesivas.
Bacterial Mutations

–Spontaneous mutations

–Induced mutations

–DNA repair mechanism

–Transposable genetic
elements

–Ames test for mutagens

 Induced Mutations

•Ultraviolet light(UV) induces adjacent thymine

molecules to link together.
 Induced Mutations

Nitrous acid–converts DNA adenine to hypoxanthine

Hypoxanthine complemented by C. Give AT to GC base pair change.

•Base analogs–substances bearing a chemical resemblance to nitrogenous

bases.

Example: 5-bromouracil
–Base analogs can inhibit DNA replication. Acyclovir,a drug against
Herpes viruses, is a base analog.
 TYPES OF MUTATIONS

•1. Base-pair change.

•2. Deletion–loss of DNA

•3. Insertion–gain of DNA

–Insertions-deletions can be cause by agent like

benzopyrene (smoke) and aflatoxin (made by
fungi).

–Insertions-deletions cause Frameshift mutations.

 Mutaciones: Desplazamiento del marco de lectura

El código genético se lee en tripletes, hay 3 modos posibles de traducir a

proteínas una secuencia de nucleótidos (dependiendo el punto de
iniciación), a esto se le llama marco de lectura. Si tenemos la secuencia
ACGACGACGACGACGACG

Las 3 marcos de lectura serán

ACG ACG ACG ACG ACG ACG
CGA CGA CGA CGA CGA CGA
GAC GAC GAC GAC GAC GAC
 DNA Repair Enzymes

•DNA repair enzymes locate and repair alteration ( mismatched

bases) and distortions (thymine dimers) of the DNA

–Mismatch repair –first proof-reading by DNA polymerase as it

is synthesizing.

–Damage/excision repair –nucleases that cut out of damaged

DNA which is repaired by DNA polymerases
Excision repair
Transposable genetic elements

•Insertion sequences

•Transposons (jumping genes)

 Transposable genetic elements

•Insertion sequences–small segments of DNA about 1000 base

pairs

–That forms a copy of itself and

–Can move (trans-locate) elsewhere on the chromosome.

–Cause of some spontaneous mutations

Transposable genetic elements

•Transposons–Larger than insertion sequences

and often carry drug resistant genes.

–Transposons move from

•Plasmid to plasmid
•Plasmid to chromosome
•Chromosome to plasmid

–End of transposons have inverted repeat

sequences.
A is the reverse and complement of the base sequence in B. (Inverted
repeat)
Ames MutagenTest

•Purpose to look for

revertant mutants

•of a histidine requiring

strain

•on media without

Histidine.
 Mutaciones rho

Las mutaciones rho muestran amplias variaciones en su influencia

sobre la terminación. La naturaleza de los efectos es la falta de
terminación.

Algunas mutaciones rho puede ser suprimidas por mutaciones en otros

genes. Las mutaciones en el gen rpoB pueden reducir la terminación
en un sitio rho dependiente.
 LexA normally binds to the
promoter regions and
represses more than 40 genes
lexA
DNA OFF =RecA
replication
bubble umuC uvrA recA
=LexA
OFF OFF OFF

However, following UV-

induced DNA damage or
lexA other replication arresting
events...
OFF
=UV-induced
umuC uvrA recA DNA damage
OFF OFF OFF

lexA ...RecA binds to the single

strand regions that are
OFF generated when replication
forks are arrested
umuC uvrA recA
OFF OFF OFF
The bound RecA serves to
protect the replication fork DNA
from degradation and also
lexA promotes the autocatalyic
ON cleavage and subsequent
degradation of the LexA
umuC uvrA recA protein, leading to the
derepression of the SOS
ON ON ON genes.
Once the arresting lesions have
been repaired and replication
has been restored, the single
lexA strand substrate for RecA
binding is no longer present,
OFF LexA is no longer cleaved and
its cellular concentration
umuC uvrA recA accumulates. Then, once again,
LexA binds to the promoters
OFF OFF OFF and represses the expression
of the SOS genes, including
that of lexA and recA.