Bact 303

Bacterial Genetics I

The Central Dogma:

DNA replication, Transcription
and Translation

The Central Dogma of Molecular Biology:

DNA------>RNA------>protein
The central dogma concerns the flow of biological
information: DNA is a self-replicating molecule containing
genetic information that can be transcribed into an RNA
message that can be translated into a polypeptide (protein).

The Central Dogma of molecular biology

Synthesis of three types of informational molecules: DNA,

RNA and Protein. Note that only one strand of the DNA is
transcribed into RNA.

In procaryotes, transcription and translation are

coupled, meaning that as an RNA message is
being transcribed, ribosomal binding to the mRNA
and the intiation of translation can begin.

Transcription and translation in a procaryote

A single mRNA often contains more than one coding region,

separated during translation into distinct polypeptides.
Also, in procaryotes transcription and translation are coupled,
meaning that they occur simultaneously.

Transcription and translation in a eucaryote

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Noncoding regions of mRNA (called introns) must be removed before

translation, leaving exons to be joined together to form a mature mRNA.
Transcription and translation cannot be coupled because transcription
takes place in the nucleus and translation occurs in the cytoplasm.

A gene is a segment of DNA that encodes for (specifies) a

protein (via mRNA), a tRNA or an rRNA.
A genome refers to the total complement of DNA in a cell.

Flow of genetic information:

DNA replication
The DNA molecule is a double
helix of two long chains of
nucleotide molecules. During
DNA replication DNA is
duplicated, producing two double
helices.

Flow of Genetic information: Transcription

Transfer of the information to RNA is called transcription,
and the molecule that encodes for one or more polypeptides
(proteins) is called messenger RNA (mRNA).

Flow of Genetic information: Translation

During translation, this genetic code in mRNA is read and
converted into protein by means of the protein
synthesizing machinery, which consists of ribosomes,
tRNA, amino acids, and a number of enzymes.

The Structure of DNA

Four nucleic acid bases are found in

DNA, adenine (A), guanine (G),
cytosine (C) and thymine (T)

The Structure of DNA

The double helix of DNA has a complementary and antiparallel

structure. One chain ends in a 5'- phosphate and the opposite
chain ends in a 3'-hydroxyl.

The Structure of DNA

DNA replication is semiconservative

DNA replication
Is bidirectional

The replication fork where the action takes

place.
During replication, as the
DNA is unwound and the
two strands are
separated, the newly
synthesized strand that
is oriented 5'-->3' is
called the leading
strand, and the new
strand that is oriented
3'-->5' is called the
lagging strand.

Transcription Fundamentals
Transcription is synthesis of RNA using a DNA template
Three Key differences between DNA and RNA:
1. RNA contains the sugar ribose instead of deoxyribose
2. RNA contains the base uracil instead of thymine
3. Except in certain viruses RNA is not a doublestranded molecule
Three major types of RNA - all products of transcription of DNA:
1. messenger RNA (mRNA)
2. transfer RNA (tRNA)
3. ribosomal (rRNA)

Transcription is the process by which nucleotide

sequences in DNA are copied to a complementary
copy of messenger RNA

RNA polymerase is the

enzyme that copies DNA into
a complementary copy of
RNA.
The enzyme uses DNA as a
template and since it
catalyzes the addition of
ribonucleotides in 5'-->3'
direction, it reads its template
DNA in the 3'-->5' direction.

Promoters
Promoters are specific sites
on DNA that RNA polymerase
first binds to to initiate the
transcription of a gene.

Sigma factors
Sigma factors are one
component of the
multicomponent RNA
polymerase enzyme that
allow the RNA polymerase to
recognize the initiation
(promoter) site.

Transcription Terminators
Transcription terminators are
sequences of nucleotide
bases at the end of the gene
that signal termination of
transcription.

During translation, the genetic code in mRNA is read

and converted into protein by means of the protein
synthesizing machinery, which consists of ribosomes,
tRNA, amino acids, and a number of enzymes.

The Genetic Code

allows for correspondence between triplets of bases in DNA and the
amino acid sequence of a polypeptide (protein)
written or expressed in terms of RNA triplets as compared to DNA triplets
because it is with messenger RNA that the translation process occurs

The Genetic Code

Codons: Triplets of three bases in RNA that encode an amino acid. There are 64 possible
codons (4 bases taken 3 at a time = 43)
Stop and start codons:
Start = AUG (codes for methionine) - site where translation begins
Stop = UAA, UAG and UGA - sites where translation ends
Degeneracy: Most amino acids have more than one codon. For example, glycine is
encoded by GGU GGC GGA and GGG.

The structure of transfer RNA (tRNA)

The bacterial ribosome

Steps in Protein Synthesis

1. initiation
begins with an initiation complex
made up ribosomes, initiation factors,
mRNA and formyl methionine tRNA.
The formyl methionine is delivered to
the initiation site on the mRNA, codon
AUG.
There is a specific sequence of bases
on the mRNA, upstream of the start
codon, that allows the ribosome to
recognize and bind onto the start site to
initiate the translation process.

Steps in Protein Synthesis

2a. elongation
The mRNA is threaded through the
ribosome which contains other sites
where tRNAs interact. The acceptor
(A) site is the site where the new
charged tRNA first attaches. The
peptide (P) site is the site where a
growing peptide is held by a tRNA and
where peptide bond formation takes
place between the incoming amino acid
(at the A site) and the amino acid at the
P site.

Steps in Protein Synthesis

2b. translocation
the ribosome advances by three
nucleotides exposing a new codon at
the A site and pushing the now empty
tRNA to the exit (E) site where it is
released from the ribosome.

Steps in Protein Synthesis

3. termination
occurs when a nonsense (stop) codon
is encountered. No tRNA binds to a
stop codon. Instead, specific proteins
called release factors recognize this
chain terminating signal and cleave the
completed polypeptide from the
terminal tRNA.

Overview of Translation

Role of ribosomal RNA in protein

synthesis
Ribosomal RNA plays a key role
at all steps of protein synthesis, in
recognition of initiation sequences,
through rRNA-mRNA interactions
during elongation, through
stabilization of tRNAs, and in a
catalytic way during peptide bond
formation and the termination
reactions.

Effects of antibiotics on protein

synthesis
A large number of antibiotics
inhibit protein synthesis in bacteria
by interacting with the ribosome.
These reactions are quite specific
and many have been shown to
interact with rRNA. Several of
these antibiotics are clinically
useful, including streptomycin,
which inhibits initiation, and
chloramphenicol and tetracycline,
which inhibit chain elongation.