BACCALAUREATE 2008 - Pearson Education...3 DNA REPLICATION 3.4 DNA replication Assessment statements 3.4.1 Explain DNA replication in terms of unwinding the double helix and separation

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DNA REPLICATION3

DNA replication3.4

Assessment statements

3.4.1 Explain DNA replication in terms of unwinding the double helix and separation of the strands by helicase, followed by formation of thenew complementary strands by DNA polymerase.

3.4.2 Explain the signifi cance of complementary base pairing in theconservation of the base sequence of DNA.

3.4.3 State that DNA replication is semiconservative.

Interesting fact

Helicase may catalyse the

unzipping of DNA at a rate

measured in hundreds of base

pairs per second.

DNA replication involves ‘unzipping’ Cells must prepare for a cell division by doubling the DNA content of the cell in a process called DNA replication. This process doubles the quantity of DNAand also ensures that there is an exact copy of each DNA molecule. You shouldtry to picture the environment in which the DNA is actually replicating. This is the environment of the nucleus during interphase of the cell cycle. Duringinterphase, there is a nuclear membrane which separates the fl uid of the nucleus(nucleoplasm) from the cytoplasm. The DNA is in the form of chromatin (nottightly coiled chromosomes). Among the variety of molecules present in thenucleoplasm there are two types that are particularly important for the process of DNA replication; they are:

enzymes needed for replication – these include helicase and a group of enzymes collectively called DNA polymerase;free nucleotides – these are nucleotides that are not yet bonded and are foundfl oating freely in the nucleoplasm, some contain adenine, some thymine,some cytosine and some guanine. (Free nucleotides are more properly called nucleoside triphosphates.)

Key fact

DNA must replicate before any

cell divides. This means that DNA

replication precedes binary fi ssion

for prokayotes, mitosis for many

eukaryotic cells and meiosis for

eukaryotic gamete and spore

producing cells.

One of the early events of DNA replication is the separation of the double helix into two single strands. DNA will lose its characteristic double helix shape when beginning this process.You should remember that the double helix is held together by the hydrogen bonds between complementary base pairs (A and T, C and G). The enzyme that initiates this separationinto two single strands is called helicase. Helicase begins ata point in or at the end of a DNA molecule and moves onecomplementary base pair at a time, breaking the hydrogenbonds so the double-stranded DNA molecule becomes twoseparate strands.

The unpaired nucleotides on each of these single strands can now be used as a template to help create two double-strandedDNA molecules identical to the original. Some people usethe analogy of a zipper for this process. When you pull on a zipper, the slide mechanism is like helicase. The separation of the two sides of the DNA molecule are like the two openedsides of a zipper (see Figure 3.13).

Helicase (currently at about the half-

way point in this image) would have

started on the left and would be moving

towards the right.

Weblink

The experimental work which

determined that DNA replication

was semiconservative is often called

‘the most beautiful experiment

in biology’. See how this classic

experiment was conducted at

http://users.rcn.com/jkimball.

ma.ultranet/BiologyPages/M/

Meselson_Stahl.htmlFormation of two complementary strandsAs shown in Figure 3.13, once DNA has become ‘unzipped’, the nitrogenousbases on each of the single strands are unpaired. In the environment of thenucleoplasm, there are many free-fl oating nucleotides. These nucleotidesare available to form complementary pairs with the single-stranded nucleotides of the unzipped molecule. This does not happen in a randomfashion. A free nucleotide locates on one opened strand at one end andthen a second nucleotide can come in to join the fi rst. This will requirethat these two nucleotides become covalently bonded together as they are the beginning of a new strand. The formation of a covalent bond betweentwo adjoining nucleotides is catalysed by one of the DNA polymerase enzymes that is important in this process.

A third nucleotide then joins the fi rst two and the process continues in a repetitive way for many nucleotides. The other unzipped strand alsoacts as a template for the formation of another new strand. This strandforms in a similar fashion, but in the opposite direction to the fi rst strand.Notice that one strand is replicating in the same direction as helicase ismoving and the other strand is replicating in the opposite direction.

The pattern of DNA replication ensures that two identical copies of DNA areproduced from one. Figure 3.14 illustrates a very small section of DNA replicating.

TOK

Who should decide how fast and

how far humans should go with our

study of DNA and the technology

that is rapidly emerging?

Notice that in the area where replication has already taken place, the two strands are absolutely identical to each other. This is because the original double–stranded molecule had complementary pairs of nucleotides and it was the complementary nucleotidesthat used the unzipped single-stranded areas astemplates.This also means that no DNA molecule is evercompletely new. Every DNA molecule after replicationconsists of a strand that was ‘old’ now paired with a strand that is ‘new’. DNA replication is described as a semiconservative process because half of a pre-existingDNA molecule is always conserved (saved).

7 The concept of semiconservative DNA replication has some interesting repercussions. For

example, one can argue that there never is such a thing as a ‘new’ DNA molecule. How long

has your DNA been in you? In your family lineage?

8 Most DNA mutations occur during DNA replication. Suggest how a mutation called a

deletion could occur. Suggest how a mutation called a substitution could occur.

A small section of DNA (shown in the

centre) is seen in a DNA polymerase

enzyme.

C A G G T A G T A C C T A C G G T A T C G

C A T G G A T G C C A T A G C

G T C C A T

helicase moving

unzipped section

Figure 3.13 The fi rst step of

DNA replication is helicase

unzipping the double-

stranded DNA molecule

forming a section with two

single strands.

C A C T A C G A T T C G C C T T A G G A C C T A C C T C A C

G G AA T C C T G G

A T G GA G T G

C T A A G C G G A A T C C T G G A T G G A G T G

T C G C C T T A G G A C C T A C C

G T G A T G

helicase

DNA polymerase

DNA polymerase

direction of synthesis

direction of synthesis

PG

PC

PC

PT

Figure 3.14 DNA replication

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BACCALAUREATE 2008 - Pearson Education...3 DNA REPLICATION 3.4 DNA replication Assessment statements 3.4.1 Explain DNA replication in terms of unwinding the double helix and separation