:

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

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

Learning Outcomes : 6.2 Describe semi-conservative replication of DNA

Models of DNA replication

Conservative
Semi-conservative Dispersive

Learning Outcomes : 6.2 Describe semi-conservative replication of DNA

Conservative
The parental double helix remains intact and an all new copy is made

Learning Outcomes : 6.2 Describe semi-conservative replication of DNA

Semi-conservative
The two strands of the parental molecule separate, and each function as a template for synthesis of a new complementary strand

Learning Outcomes : 6.2 Describe semi-conservative replication of DNA

Dispersive
Each strand of both daughter molecule contains a mixture of old and newly synthesized parts

Learning Outcomes : 6.2 Describe semi-conservative replication of DNA

Evidence for semi-conservative hypothesis

Experiment performed by Meselson & Stahl in 1950, using the radioisotope labelling technique 2 different isotopes of Nitrogen are used : a) 14N is the light isotope of nitrogen b) 15N is the heavy isotope of nitrogen

Fig. 14.12b

Learning Outcomes : 6.2 Describe semi-conservative replication of DNA

Conclusion:
The results indicates that, replication of DNA strands by mean of semi-conservative mechanism
Each strand of a DNA double helix acts as a template for the synthesis of a new strand

Learning Outcomes : 6.2 Describe DNA replication during cell division

Overview of DNA replication

Learning Outcomes : 6.2 Describe DNA replication during cell division

Origin of replication
A special sites where the replication of DNA molecule begins

It is where two parental strands separate and form replication bubbles

Learning Outcomes : 6.2 Describe DNA replication during cell division

In bacterial chromosome (circular DNA) has a single origin

In eukaryotes, DNA replication begins at many sites along the giant DNA molecule of each chromosome

Learning Outcomes : 6.2 Describe DNA replication during cell division

Replication process The steps:

1. DNA gyrase relieves strain of double-stranded DNA 2. A portion of the parental double helix is unwound by a DNA helicase.

Learning Outcomes : 6.2 Describe DNA replication during cell division

3. Molecules of single strand binding protein stabilize the unwound template strands 4. RNA primer is first synthesized at the origin of replication

Learning Outcomes : 6.2 Describe DNA replication during cell division

5. RNA primer is synthesized by a protein complex known as RNA primase 6.Primers are generally 5-10 nucleotides long

Learning Outcomes : 6.2 Describe DNA replication during cell division

7. RNA primase removed, DNA polymerase III synthesized DNA nucleotides to RNA primer

Learning Outcomes : 6.2 Describe DNA replication during cell division

8. DNA polymerase III add nucleotides only to the free 3 end of growing DNA strand, never to the 5

9. Thus, a new DNA strand can elongate only in the 5 to 3 direction

Learning Outcomes : 6.2 Describe DNA replication during cell division

10. DNA polymerase I then replace the RNA primer with DNA nucleotides
11. Finally DNA ligase joins the 3 end of the DNA that replaces the primer to the rest of new strand

Learning Outcomes : 6.2 Describe DNA replication during cell division

Leading and Lagging strands

Along one template strand, DNA polymerase III adding nucleotides in 5 to 3 direction continuously, towards the replication fork

The DNA strand made by this mechanism is called leading strand

Learning Outcomes : 6.2 Describe DNA replication during cell division

To elongate the other new strand of DNA, DNA polymerase III must work along the other template strand in the direction away from the replication fork

The DNA strand synthesized in this direction is called lagging strand

Learning Outcomes : 6.2 Describe DNA replication during cell division

In the lagging strand, only short piece of DNA can be synthesized because DNA polymerase III need to move far away from the fork This short segment is called Okazaki fragments Each Okazaki fragment initiated by separate RNA primer

Learning Outcomes : 6.2 Describe DNA replication during cell division

DNA polymerase III adds DNA nucleotides to the primer forming Okazaki fragment After reaching the next RNA primer, DNA polymerase III falls off DNA polymerase I replaces the RNA primer with DNA nucleotide Finally, DNA ligase joins the fragments of the lagging strand together