1. the amount of A, T, G, C in DNA varies
from specie to specie
2. in each species the amount of A=T
and the amount of G=C
Structure
COMPLEMENTARY PAIRING: A is always bonded to
T & G is always bonded to C by a hydrogen bond
A purine (wide) is always paired to a
pyrimidine (narrow) to complement sizes
Strands with ANTI PARALLEL ARRANGEMENT
Every nucleotide has a phosphate
group at the 5' position of the sugar
& they bind together by linking this 5'
phosphate to the free hydroxyl (--OH)
at the 3' position of the other
nucleotide's sugar (giving it it's
direction)
DOGMA OF MOLECULAR BIOLOGY
Replication: Copying a DNA molecule
DNA replication is termed
semi-conservative replication because
each daughter DNA double helix
contains an old strand from the parental
DNA double helix and a new strand
STEPS:
1. Unwinding: an enzyme called helicase breaks the hydrogen bond
between the paired bases (creating a replication fork)
2. Complementary Base Pairing: New complementary nucleotides, always present in the
nucleus, are positioned in place (by an enzyme complex called DNA polymerase)
3. Joining: new strands are crated so each daughter DNA molecule contains
an old and a new strand (by an enzyme complex called DNA polymerase)
DNA ligase, joins the fragments, but there is
no way for DNA polymerase to replicate the
5' ends of both new strands after RNA
primers are removed. So DNA molecules get
shorter as one replication follows another.
the leading new strand continues it's
normal process, the lagging strand is
discontinuous, the segments are known
as Okazaki Fragments. as the RNA
primers are replaced by the nucleotides.
A DNA polymerase is very
accurate and makes a mistake
approximately once per
100,000 base pairs at most.
Differences
between...
Eukaryotes
Telomeres (special
nucleotide sequence at the
end of the DNA) do not code
for proteins, instead creates
repeats of a short nucleotide
sequence, such as TTAGGG.
replication may begin at numerous origins
the process can take
hours because of the
amount of info replicated
Prokaryotes
Bacteria have a single
circular loop of DNA,
replication moves around
the DNA molecule in one
direction only.
it requires 40 min. to
replicate & can do so
every 20 min.
it is possible for a
new round of DNA
replication to begin
even before the
previous round is
complete.
Transcription
1st step in syntetization of a protein: DNA
serves as a template for RNA formation.
pre-mRNA if formed (by RNA
Polymerase when it ataches to
a DNA promoter)
RNA (Ribonucleic Acid)
carries the information
Polymere composed of nucleotides:
sugar ribose & bases.
Structure: single stranded,
NO double helix
typically has a poly a tail (has many adenine
nucleotides at one end & a cap at the other end)
Messenger RNA (mRNA) takes a
message from DNA in the nucleus to
the ribosomes in the cytoplasm.
Transfer RNA (tRNA) transfers
amino acids to the ribosomes
Ribosomal RNA (rRNA), along with ribosomal
proteins, makes up the ribosomes, where
polypeptides are synthesized.
Codon (triplet code): 3
nucleotide bases
properties
1. The genetic code is
degenerate. most amino
acids have more than
one codon; (leucine,
serine, and arginine have
six different codons)
The degeneracy of the code
protects against potentially
harmful effects of mutations.
2. The genetic code is unambiguous. Each
triplet codon has only one meaning.
3. The code has 1 start
and 3 stop signals.
elongation of the mRNA
continues until DNA
reaches a stop point & the
mRNA is liberated = mRNA
Transcript
made up of nucleotides & regions
called: exons and introns
splicing: introns are removed (forming
an mRNA: template to form proteins)
m RNA will leave the nucleus
through the nuclear pore into
the cell's cytoplasm
Prokatyotes have
self-spllicing: the
intron itself has
the capability of
enzymatically
splicing itself out
of a premRNA.
Another type of RNA
called small nucleolar
RNA (snoRNA) is present
in the nucleolus, where
it helps process rRNA and
tRNA molecules.
Eukaryotes: splicing is
done by spliceosomes,
which contain small
nuclear RNAs (snRNAs:
are capable of
identifying the introns to
be removed) A
spliceosome utilizes a
ribozyme when it
removes the introns.
REGULATION OF GENE ACTIVITY
Prokaryote
Regulation
Operon
Model
Promoter: signals where
transcription is to begin.
Operator: controls transcription of
structural genes.
active repressor
binds to the
operator, RNA
polymerase
cannot attach to
the promoter, and
transcription
cannot occur
Structural Genes: instructions to primary
structure of enzymes in a metabolic pathway
Regulator Gene: codes for a repressor that
controls whether the operon is active or not.
Trp Operon
(repressible system)
when the active repressor
binds to the operator
transcription is prevented
because RNA polymerase
can't bind to the promoter
gene expression "on" by default the
corepressor can bind to the repressor
activating it causing expresion to go
off
only on/off
(negative gene
regulation)
Lac Operon (inducible
systems)
on/ off
(negative
gene
regulation)
gene expression "off" by
default corepressor binds to
repressor deactivating it and
turnig expresion "on" by
unbinding it form the operator
by unbinding the repressor
from the promoter
expression is allowed
up/ down
(positive gene
regulation)
there are no
repressors,
just
activators
there's some
level of
transcription
but
we
need
more
cyclic amp
activates the
activator by
binding to it
and changing
it's form so it
can bind with
the promoter
and increase
transctription
Eukaryote Regulation
1. Chromatin Structure
core of 8 histone
proteins wraped around
by DNA
the DNA packed
around the histone
can't be transcribed
chromatin remodeling
complex can liberate the DNA
making it transcribable
Heterochromatin:
strongly
packed
histones
Euchromatin:
loosely packed
histones
Epigenetic inheritance: When
histones are methylated, sometimes
the DNA itself becomes methylated
as well = genetic imprinting,
expression depends if the gene is
inherited from the mother or father,
not the gene it self
2. Transcriptional Control
transcription
factors: proteins
that help regulate
transcription
transcription
activators are DNA
binding proteins that
speed transcription
dramatically.
transcription factors bind to the
promoter and transcription
activators bind to an enhancer
when the loop is formed to allow
transcription to begin
3. Posttranscriptional Control
occurs in the nucleus
and includes alternative
mRNA splicing and
controlling the speed
with which mRNA
leaves the nucleus.
pre-mRNA has introns
and exons, introns stay in
the nucleus as exons exit
the nucleous = mRNA
4. Translational
Control
mRNA strands after
splicing are too short
so they become...
micro RNA's, they
are double stranded
and where turned
off, micro RNA can
lower the expression
of genes
5. Posttranslational Control
the last chance a cell
has for influencing
gene expression
Some proteins are
not immediately
active after
synthesis.
some proteins only
become active when it
is appropriate for them
to do so.
Gene
Mutations
permanent change
in the sequence of
bases in DNA
Causes
Spontaneous mutations
the movement of
transposons from
one chromosomal
location to another
can disrupt a gene
and lead to an
abnormal product.
a base in DNA can undergo a chemical
change that leads to a miss pairing
during replication and may be carried
in future generations.
they are extremely
rare, 1 billion
nucleotide pairs
replicated.
Induced
Mutations
caused by mutagens,
environmental factors that can
alter the base composition of DNA
Many mutagens
are also
carcinogens
(cancer-causing)
industrial
chemicals, foods,
tabacco, radiation
Effects
on Protein
Activity
point mutation: e a change in a single
DNA nucleo - tide and, therefore, a
possible change in a specific amino acid.
can range in effect,
depending on the
particular codon
change
Frameshift mutation:
one or more nucleotides
are either inserted or
deleted from DNA.
The result can be a completely new sequence
of codons and nonfunctional protein
Non-Functional Proteins
dramatic effect on the
phenotype, because
enzymes are often a part
of metabolic pathways.
Mental retardation,
albinism, PKU, etc.
Cancer
The development of
cancer involves a
series of accumulating
mutations that can be
different for each type
of cancer
tumor suppressor
genes are inactive
and oncogenes are
active
cell division occurs
uncontrollably because a cell
signaling pathway that reaches
from the plasma membrane to
the nucleus no longer
functions as it should
Translation
2nd step in syntetization of a
protein: the mRNA transcript
directs the sequence of amino
acids in a polypeptide.
cell changes a nucleotide
sequence into an amino acid
sequence. With the help of
the three types of RNA, a
gene (a segment of DNA)
specifies the sequence of
amino acids in a polypeptide
A tRNA molecule is a single-stranded nucleic
acid that doubles back on itself to create
regions where complementary bases are
hydrogen-bonded to one another
there's at least 1 for every amino acid in the
protein, it binds to the 3' end.
on the 5' end, an anti codon (group of three
bases complementary to a specific mRNA codon
so that they pair in an antiparallel fashion)
fewer tRNAs that
codos because of the
wobble hypothesis.
the first two positions in a tRNA anticodon
pair obey the A–U/G–C configuration, but
the third position can be variable. This
helps ensure that despite changes in DNA
base sequences, the correct sequence of
amino acids will result in a protein
aminoacyl-tRNA synthetase (enzyme found in the
cytoplasm) attaches a different type of aminoacid to the
specific tRNA
this process uses
ATP
amino acid–tRNA complex is formed
travels to a ribosome
formation in eukaryotes: rRNA
spliced to form 2 rRNA strtands, 1
small & 1 big
produced from a DNA template in the nucleolus
of a nucleus, synthetised by RNA polymerase I
they leave the nucleolus & fold to form a
small and large sub unit of the ribosome
= pre-ribosome (still inside the nucleus)
they leave the
nucleous and the
ribosome can
either stay un the
cytoplasm or get
attatchd to the ER
the small and large sub units of the
ribosome attach to the cap of the mRNA
Translation
begins
STAGE 1 Initiation: Proteins (initiation factors)
assemble the small ribosomal subunit and the
large ribosomal subunit through the P site of the
ribosome, now occupied by the initiator tRNA (with
an attached peptide), to the cap of the mRNA
In prokaryotes, a small
ribosomal subunit attaches
to the mRNA in the vicinity of
the start codon (AUG).The
first or initiator tRNA pairs
with this codon. Then, a large
ribosomal subunit joins to
the small subunit
STAGE 2 Elongation:
1 tRNA + amino acid
arrives at the A site.
2 ribosome verifies that
new tRNA matches the
codon and firmly places it
at the A site, peptide will
transfer to the new tRNA.
3 the peptide bond
formation is one amino
acid longer than it was
before. creating a protein.
(now attached to the new
tRNA at A site)
4 translocation occurs: The ribosome
moves forward, and the peptide-bearing
tRNA is now at the P site of the ribosome.
The spent tRNA is now at the E site, and it
exits. :A new codon is at the A site and is
ready to receive another tRNA. (this
process repeats until the end of mRNA
strand)
STAGE 3 Termination: it occurs at a
stop codon (codon that don't code
for an amino acid), a protein
called a release factor, binded to a
stop codon, releases the tRNA at P
site and the polypeptide chain.
the ribosome separates into 2,
structure: ribosome
has three binding
sites for tRNAs.
E (exit) site
P (peptide) site
A (amino acid) site
DNA synthesised by RNA polymerase III creates the tRNA
The universal nature of the genetic
code provides strong evidence that
all living things share a common
evolutionary heritage
. Since the same genetic code is used by
all living things, it is possible to transfer
genes from one organism to another.