SCORE A CLASS

CHAPTER 6

EXPRESSION OF BIOLOGICAL INFORMATION

6.0 Expression of Biological Information

6.1 DNA and genetic information 6.2 DNA replication 6.3 Protein synthesis: transcription and translation 6.4 Gene regulation and expression lac operon

Structure of DNA

Scientist and their experiment

Scientist Watson and Crick Griffith (1928) Material /organism use In experiment DNA Streptococcus pneumoniae (S strain and R strain) and mice Streptococcus pneumoniae (S strain and R strain), mice and enzyme protease, Rnase and Dnase. Bacteriophage and E. coli Conclusion Double helix of DNA Discovery of transforming agent in bacteria. DNA is transforming agent

Avery et. al. (1944)

Hershey & Chase (1952)

DNA is genetic material of viruses

Beadle and Tatum (1941)

Meselson and Stahl (1950)

Bread mole, Neurospora crassa

E.Coli and radioactive isotop N14 and N15

One gene, one polypeptide

DNA replication occur in semiconservative way

DNA REPLICATION

Overview

DNA DNA Replication Replication

Single-strand binding protein Helicase

Origin of replication Leading strand Lagging strand

Lagging strand

Leading strand

Overall directions of replication

Leading strand

5 3

DNA pol III Primer Primase 3 DNA pol III 5

4

Parental DNA

Lagging strand DNA pol I 5

3
2

DNA ligase 5
1

Enzyme involve in DNA replication

Protein Helicase Function for Leading and Lagging Strands Unwind parental double helix at replication forks

Singlestrand binding protein Topoisomerase (gyrase)

Binds to and stabilizes single-stranded DNA until it can be used as a template

Corrects 'overwinding' ahead of replication forks by breaking, swivelling and rejoining DNA strands

Enzyme involve in DNA replication Enzyme involve in DNA replication

Protein Function for Leading Function for Lagging strand strand
Synthesizes single RNA primer at the 5' end of the leading strand Continously synthesizes the leading strand, adding on to the primer Synthesizes single RNA primer at the 5' end of each Okazaki fragment Elongates each Okazaki fragment, adding on to its primer

Primase

DNA Polymerase III

Enzyme involve in DNA replication

Protein Function for Leading strand Function for Lagging strand

DNA Polymerase I

Removes primer a leading strand and replace it with DNA nucleotide, adding on to the adjacent 3' end

Removes primer from each Okazaki fragment and replace it with DNA nucleotide, adding on to the 3' end of adjacent fragment
Joining the sugarphosphate backbones of all the Okazaki fragments into a continuous DNA strand

DNA Ligase

PROTEIN SYNTHESIS - Transcription -Translation

The roles of transcription and translation

From Gene to

Protein (e.g. : enzymes)

12

Eukaryotic Cell

Prokaryotic Cell

The nucleus provides a separate compartment for transcription. The original RNA transcript, called pre-mRNA, is process in various ways before leaving the nucleus as mRNA.

In a bacterial cell, which lacks of nucleus, mRNA produced by transcription is immediately translated without additional processing.

Transcription
Transcription can be divide into three stages i. Initiation ii. Elongation iii. Termination

Copyright 2002 Pearson Education, Inc., publishing as Benjamin Cummings

14

Overview of Transcription

mRNA processing

In RNA processing: Introns (non-coding region) are removed and Exons (coding region) are spliced/joined together to create mature mRNA molecule with a continuous coding sequence.

Differences between DNA replication and transcription

DNA replication The whole length of DNA is used Produced double stranded DNA Two stranded of DNA are used as template Primer is require DNA polymerase is used Transcription Only portion of DNA is used Produced single stranded mRNA One stranded of DNA is used as template Primer is not require RNA polymerase is used

Genetic code

Genetic code is code which specifies the relationship between the sequence of nucleotides in the mRNA and the sequence of amino acid in a polypeptide

Genetic code: 64 codons

Translation

Molecular component in translation

1) mRNA

2) rRNA (Ribosome)
3) tRNA with amino acid

tRNA structure

Activation of amino acids for attachment to tRNA

Schematic model with mRNA and tRNA

Growing polypeptide Amino end Next amino acid to be added to polypeptide chain

tRNA 3

mRNA

Codons

25

3 steps of translation process; synthesis of a polypeptide chain

1) Initiation 2) Elongation - Codon recognition - Peptide bond formation - Translocation

3) Termination

Initiation

2 Elongation
The translocation step requires energy, which is provided by hydrolysis of GTP molecules.

3.

Termination

Polyribosome/Polysomes
Multiple ribosomes can translate a single mRNA
at the same time, forming a polyribosome (or polysome) Thus making it possible to produce many polypeptides simultaneously from a single mRNA at one time

Lac Operon
What is Operon?

An operon is cluster of
bacterial genes along with an adjacent promoter that controls the transcription of those genes.

The bacterial genes on DNA

Regulatory gene
lac I
lac Operon Gene
P O Lac Z

Lac Operon
P O lac Z lac Y lac A
Gene label
Promoter : site where RNA polymerase bind Operator : Site where repressor protein bind Gene for -galactosidase

Lac Y
Lac A

Gene for Lactose permease

Gene for transacetylase

Lactose absent

Lactose present

How to memorize easily.

RP
Repressor protein Lac Y

Regulatory gene

Lac I

O
Operator

Promoter

Lac Z

Lac A

Figure 1: Lac operon structure

How to memorize easily.

RP

Lac I

Figure 2: When lactose is present. Finger show GOOD sign RP will NOT bind at O

Lac operon is ON

How to memorize easily.

RP

Lac I

- galactosidase

Figure 3: Transcription occur Lac Z produce - galactosidase

How to memorize easily.

RP

Lac I

- galactosidase Lactose permease

Figure 4: Transcription occur Lac Y produce lactose permease

How to memorize easily.

RP

Lac I

- galactosidase Lactose permease transacetylase

Figure 5: Transcription occur Lac A produce transacetylase

How to memorize easily.

RP

Lac I

Figure 6: Lactose is absent.

Lac A produce transacetylase RP will bind at O. Transcription will NOT occur. Enzymes will NOT produce.