Learning Outcomes
- Identify the major enzymes that play a role in DNA replication
The process of DNA replication is catalyzed by a type of enzyme called DNA polymerase (poly meaning many, mer meaning pieces, and –ase meaning enzyme; so an enzyme that attaches many pieces of DNA). During replication, the two DNA strands separate at multiple points along the length of the chromosome. These locations are called origins of replication because replication begins at these points. Observe Figure 1: the double helix of the original DNA molecule separates (blue) and new strands are made to match the separated strands. The result will be two DNA molecules, each containing an old and a new strand. Therefore, DNA replication is called semiconservative. The term semiconservative refers to the fact that half of the original molecule (one of the two strands in the double helix) is “conserved” in the new molecule. The original strand is referred to as the template strand because it provides the information, or template, for the newly synthesized strand.
DNA polymerase needs an “anchor” to start adding nucleotides: a short sequence of DNA or RNA that is complementary to the template strand will work to provide a free 3′ end. This sequence is called a primer (Figure 2).
How does DNA polymerase know in what order to add nucleotides? Specific base pairing in DNA is the key to copying the DNA: if you know the sequence of one strand, you can use base pairing rules to build the other strand. Bases form pairs (base pairs) in a very specific way.
Practice Questions
True/False: DNA replication requires an enzyme.
What are the building blocks on DNA?
- Deoxyribonucleotides
- Fatty acids
- Ribonucleotides
- Amino acids
True/False: DNA replication requires energy.
We have the building blocks, a source of energy, and a catalyst. What’s missing? We need instruction about the order of nucleotides in the new polymer. Which molecule provides these instructions?
- Protein
- DNA
- Carbohydrate
- Lipid
There is one more thing required by the DNA polymerase. It cannot just start making a DNA copy of the template strand; it needs a short piece of DNA or RNA with a free hydroxyl group in the right place to attach the nucleotides to. (Remember that synthesis always occurs in one direction—new building blocks are added to the 3′ end.) This component starts the process by giving DNA polymerase something to bind to. What might you call this short piece of nucleic acid?
- A solvent
- A primer
- A converter
- A sealant
Now that you understand the basics of DNA replication, we can add a bit of complexity. The two strands of DNA have to be temporarily separated from each other; this job is done by a special enzyme, helicase, that helps unwind and separate the DNA helices (Figure 4). Another issue is that the DNA polymerase only works in one direction along the strand (5′ to 3′), but the double-stranded DNA has two strands oriented in opposite directions. This problem is solved by synthesizing the two strands slightly differently: one new strand grows continuously, the other in bits and pieces. The leading strand grows continuously and the lagging strand is put together with short pieces called Okazaki fragments. These fragments are connected and linked together by an enzyme called DNA ligase. Short fragments of RNA are used as primers for the DNA polymerase.
Practice Questions
Which of these separates the two complementary strands of DNA?
- DNA polymerase
- helicase
- RNA primer
- single-strand binding protein
Which of these attaches complementary bases to the template strand?
- DNA polymerase
- helicase
- RNA primer
- single-strand binding protein
Which of these is later replaced with DNA bases?
- DNA polymerase
- helicase
- RNA primer
- single-strand binding protein
In Summary: Major Enzymes
Replication in eukaryotes starts at multiple origins of replication. A primer is required to initiate synthesis, which is then extended by DNA polymerase as it adds nucleotides one by one to the growing chain. The leading strand is synthesized continuously, whereas the lagging strand is synthesized in short stretches called Okazaki fragments. The RNA primers are replaced with DNA nucleotides; the DNA remains one continuous strand by linking the DNA fragments with DNA ligase.
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