DNA Replication: Review of Enzymes, Replication Bubbles & Leading and Lagging Strands

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  • 0:05 The Big Picture of DNA…
  • 0:39 Review of…
  • 3:41 RNA Primase and…
  • 4:57 Leading and Lagging Strands
  • 7:04 Summary for DNA Replication
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Lesson Transcript
Instructor
April Koch

April teaches high school science and holds a master's degree in education.

Expert Contributor
Brenda Grewe

Brenda has 25 years of experience teaching college level introductory biology and genetics. She earned her PhD in Genetics from Indiana University.

Feeling lost in the thorny details of DNA replication? This lesson provides an overview of semi-conservative replication, with a focus on putting together all of the concepts involved. We'll review the work of each enzyme on our way to completing the big picture of DNA replication.

The Big Picture of DNA Replication

The replication of DNA is a complex process that took scientists many years and lots of hard work to understand. Previously, we discussed the fine details of semi-conservative replication: how the DNA double helix is unwound and how parent DNA is copied to produce daughter strands. We learned about a cast of helpful enzymes that make all the molecular movements possible. But, understanding how all the pieces fit together can be a challenge. So, in this lesson, we'll review all the parts of DNA replication and add some new bits of information that will help to complete the picture.

Review of Replication and Essential Enzymes

In semi-conservative replication, new daughter strands are added to parent strands that split apart.
Parent and daughter DNA strands

Let's start by reminding ourselves of the basics of semi-conservative replication. We begin with the original DNA molecule, and both strands in that molecule are referred to as parent strands. The goal of DNA replication is to make a second DNA molecule, using the parent strands as a template to create two new daughter strands. The term semi-conservative refers to the fact that both parent strands are conserved, or saved, in each of the new molecules. So, in semi-conservative replication, the parent strands split apart, but each remains whole, while new daughter strands are added onto them. The end result is two DNA molecules, each containing one parent strand and one daughter strand.

Now, let's go over all the steps of DNA replication. Along the way, we'll check in with each essential enzyme and discuss how it helps in completing each step. Since the names and functions of the enzymes can get confusing, we'll make an enzyme chart before we begin. As we discuss the steps of DNA replication, we'll fill in our enzyme chart to keep all our information organized.

Enzyme chart
Enzyme Chart

The first step occurs when DNA helicase unwinds the double helix by breaking the hydrogen bonds between the parent strands of DNA. This splitting and unwinding process opens up the DNA molecule into a Y-shape, which we call the replication fork. Already we have our first enzyme, so let's fill that in. DNA helicase is the enzyme that unwinds the DNA double helix.

Before new daughter strands can be added on, the parent strands are first made ready by an RNA primer. The primer is built by the enzyme RNA primase. So, let's go to the chart: RNA primase is the enzyme that builds an RNA primer on the parent strand to initiate DNA replication.

Once the RNA primer is built, then the next enzyme, DNA polymerase, is free to do its job. DNA polymerase slides into the replication fork and positions itself behind the RNA primer. It begins to add DNA nucleotides onto each parent strand. DNA polymerase always works by starting at the 3' end of DNA and moving toward the 5' end. This means that on the leading strand, it works continuously as it follows DNA helicase, which is constantly opening the fork more and more.

But on the lagging strand, DNA polymerase works discontinuously, making Okazaki fragments in the opposite direction. So, now that we understand what DNA polymerase does, let's go back to our chart. DNA polymerase is the enzyme that matches and lays down nucleotides to build the daughter DNA strand along each parent DNA strand.

Now we're left with all these Okazaki fragments that are separate from each other, so they need to be joined together by the enzyme DNA ligase. DNA ligase binds the fragments end to end, forming a continuous daughter strand on top of the lagging parent strand. So, that was our last enzyme: DNA ligase joins the adjacent Okazaki fragments on the lagging strand of DNA.

RNA Primase and Okazaki Fragments

The lagging strand is always behind the leading strand due to discontinuous replication.
Leading and lagging strands

Let's back up for just a minute. Have you noticed a couple of elements that don't quite seem to fit? If you have, then you're on the right track. Until now, we haven't had a chance to put the concept of the RNA primer together with the concept of Okazaki fragments. In earlier lessons, we talked about how the RNA primer must be present before DNA polymerase can begin to do its job. Later, when we talked about Okazaki fragments, we mentioned that DNA polymerase has to keep going back and restart its work at the beginning of every fragment. So, you may be wondering about this. Doesn't there have to be a new RNA primer to begin every new Okazaki fragment?

Well, the answer is yes. Every time DNA polymerase restarts its work, it needs an RNA primer. Of course, in the leading strand, only one primer is needed. This primer is built upon the 3' end, the free end, before replication begins moving toward the fork. But on the lagging strand, we need an RNA primer for each Okazaki fragment! Every time DNA polymerase finishes out its track and moves back for another go, it has to have a primer waiting there to help it get started again. So, on the lagging strand, there are lots of RNA primers scattered all down the line - one for every Okazaki fragment that is to be made.

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Additional Activities

How to Build a New DNA Molecule

The activity below provides an opportunity for you to apply your understanding of what you have learned about the steps and enzymes involved in semi-conservative DNA replication. A schematic diagram of one fork of a replicating DNA molecule is shown. The diagram and activity highlight two important aspects of DNA replication: 1) the two strands of a DNA molecule are anti-parallel; that is, in the opposite direction with respect to their 5' and 3' ends, and 2) DNA polymerase moves along each parent (template) DNA strand in the 3' to 5' direction, laying down the new strands in the 5' to 3' direction; that is, anti-parallel to the parent strand.

Diagram of the Replication Fork

Brenda Grewe (public domain)

Activity

Review DNA Replication video if needed, then answer the following questions. Use the schematic diagram of DNA replication provided. The 5' and 3' ends of the parent DNA strands are labeled. The 3' end of a new strand has an arrow head. The fat arrow marks the replication fork.

1. Label the newly made leading strand of DNA, including the 5' and 3' ends. Hint: the leading strand is the one that is made continuously!

2. Label the newly made lagging strand of DNA, including the 5' and 3' ends.

3. Identify the Okazaki fragments and show where a primer would be located for each. (Make the primer a different color or thickness). Hint: each Okazaki fragment must have a primer.

4. Identify the location of a primer for leading strand synthesis. Hint: only one is needed.

5. Indicate on the diagram where each of the following enzymes would be located:

a. DNA Polymerase - makes the leading as well as the lagging strand

b. DNA Ligase - joins the pieces of the lagging strand

c. Helicase - unwinds the parent DNA strands beginning at the origin of replication and as the replication fork moves

d. Primase - makes the RNA primers to initiate new strand synthesis

6. What is the origin of replication? Where would it be in the diagram? Hint: the replication fork advances from left to right beginning at the origin.

7. Describe the order of action of the enzymes involved in replication (the enzymes are 5a-d). Hint: unwinding at the origin is the first step.

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