Skip to main content
Biology LibreTexts

8.3: DNA Replication

  1. Last updated
  2. Save as PDF
  • Page ID
    35709

    • \( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\)

    What you’ll learn to do: Explain the role of complementary base pairing in the precise replication process of DNA

    In this outcome, we’ll learn more about the precise structure of DNA and how it replicates. Watch this video for a quick introduction to this topic:

    Learning Objectives
    • Outline the basic steps in DNA replication
    • Identify the major enzymes that play a role in DNA replication
    • Identify the key proofreading processes in DNA replication

    Basics of DNA Replication

    Illustration shows the conservative, semi-conservative, and dispersive models of DNA synthesis. In the conservative model, when DNA is replicated and both newly synthesized strands are paired together. In the semi-conservative model, each newly synthesized strand pairs with a parent strand. In the dispersive model, newly synthesized DNA is interspersed with parent DNA within both DNA strands.
    Figure 1. The three suggested models of DNA replication. Grey indicates the original DNA strands, and blue indicates newly synthesized DNA.

    The elucidation of the structure of the double helix provided a hint as to how DNA divides and makes copies of itself. This model suggests that the two strands of the double helix separate during replication, and each strand serves as a template from which the new complementary strand is copied. What was not clear was how the replication took place. There were three models suggested: conservative, semi-conservative, and dispersive (see Figure 1).

    In conservative replication, the parental DNA remains together, and the newly formed daughter strands are together. The semi-conservative method suggests that each of the two parental DNA strands act as a template for new DNA to be synthesized; after replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. In the dispersive model, both copies of DNA have double-stranded segments of parental DNA and newly synthesized DNA interspersed.

    Meselson and Stahl were interested in understanding how DNA replicates. They grew E. coli for several generations in a medium containing a “heavy” isotope of nitrogen (15N) that gets incorporated into nitrogenous bases, and eventually into the DNA (Figure 2).

    Illustration shows an experiment in which E. coli was grown initially in media containing ^{15}N nucleotides. When the DNA was extracted and run in an ultracentrifuge, a band of DNA appeared low in the tube. The culture was next placed in ^{14}N medium. After one generation, all of the DNA appeared in the middle of the tube, indicating that the DNA was a mixture of half ^{14}N and half ^{15}N DNA. After two generations, half of the DNA appeared in the middle of the tube, and half appeared higher up, indicating that half the DNA contained 50% ^{15}N, and half contained ^{14}N only. In subsequent generations, more and more of the DNA appeared in the upper, ^{14}N band.
    Figure 2. Meselson and Stahl experimented with E. coli grown first in heavy nitrogen (15N) then in 14N. DNA grown in 15N (red band) is heavier than DNA grown in 14N (orange band), and sediments to a lower level in cesium chloride solution in an ultracentrifuge. When DNA grown in 15N is switched to media containing 14N, after one round of cell division the DNA sediments halfway between the 15N and 14N levels, indicating that it now contains fifty percent 14N. In subsequent cell divisions, an increasing amount of DNA contains 14N only. This data supports the semi-conservative replication model. (credit: modification of work by Mariana Ruiz Villareal)

    The E. coli culture was then shifted into medium containing 14N and allowed to grow for one generation. The cells were harvested and the DNA was isolated. The DNA was centrifuged at high speeds in an ultracentrifuge. Some cells were allowed to grow for one more life cycle in 14N and spun again. During the density gradient centrifugation, the DNA is loaded into a gradient (typically a salt such as cesium chloride or sucrose) and spun at high speeds of 50,000 to 60,000 rpm. Under these circumstances, the DNA will form a band according to its density in the gradient. DNA grown in 15N will band at a higher density position than that grown in 14N. Meselson and Stahl noted that after one generation of growth in 14N after they had been shifted from 15N, the single band observed was intermediate in position in between DNA of cells grown exclusively in 15N and 14N. This suggested either a semi-conservative or dispersive mode of replication. The DNA harvested from cells grown for two generations in 14N formed two bands: one DNA band was at the intermediate position between 15N and 14N, and the other corresponded to the band of 14N DNA. These results could only be explained if DNA replicates in a semi-conservative manner. Therefore, the other two modes were ruled out.

    During DNA replication, each of the two strands that make up the double helix serves as a template from which new strands are copied. The new strand will be complementary to the parental or “old” strand. When two daughter DNA copies are formed, they have the same sequence and are divided equally into the two daughter cells.

    Click through this tutorial on DNA replication.

    Learning Objectives

    The model for DNA replication suggests that the two strands of the double helix separate during replication, and each strand serves as a template from which the new complementary strand is copied. In conservative replication, the parental DNA is conserved, and the daughter DNA is newly synthesized. The semi-conservative method suggests that each of the two parental DNA strands acts as template for new DNA to be synthesized; after replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. The dispersive mode suggested that the two copies of the DNA would have segments of parental DNA and newly synthesized DNA. Experimental evidence showed DNA replication is semi-conservative.

    Major Enzymes

    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.

    Stylized 3' synthesis shown, no enzymes in diagram.
    Figure 3. By Madprime(Wikipedia) (DNA replication split horizontal) CC BY-SA 2.0
    Diagram of a primer moving along the template strand of DNA.
    Figure 4. Primer and Template

    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 4).

    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.

    Diagram showing the hydrogen bonds between nucleotides. Adenine is bound to thymine, and cytosine is bound to guanine.
    Figure 5. DNA chemical structure. Modification of DNA chemical structure by Madeleine Price Ball; CC-BY-SA-2.0
    Practice Questions

    True/False: DNA replication requires an enzyme.

    Show Answer

    True. Most biological reactions rely on the enzyme to speed up the reaction. In the case of DNA replication, this enzyme is DNA polymerase.

    What are the building blocks on DNA?

    1. Deoxyribonucleotides
    2. Fatty acids
    3. Ribonucleotides
    4. Amino acids
    Show Answer

    Answer a. DNA is a double helix made up of two long chains of deoxyribonucleotides.

    True/False: DNA replication requires energy.

    Show Answer

    True. Making large molecules from small subunits (anabolism) requires energy. What supplies the energy? The building blocks themselves serve as a source of energy. As they get incorporated into the DNA polymer, two phosphate groups are broken off to release energy, some of which is used for making the polymer. Deoxyribonucleotides differ from nucleotides like ATP only by one missing oxygen atom.

    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?

    1. Protein
    2. DNA
    3. Carbohydrate
    4. Lipid
    Show Answer

    Answer b. We refer to this DNA as a template. The original information stored in the order of bases will direct the synthesis of the new DNA via base pairing.

    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?

    1. A solvent
    2. A primer
    3. A converter
    4. A sealant
    Show Answer

    Answer b. A primer is used to start this process by giving DNA polymerase something to bind the new nucleotide to.

    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 6). 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.

    Diagram of both the leading and lagging strands the helicase splits the two strands and a DNA polymerase travels over both strands to create complementary strands.
    Figure 6. By Mariana Ruiz (DNA replication) Public Domain
    Practice Questions

    Which of these separates the two complementary strands of DNA?

    1. DNA polymerase
    2. helicase
    3. RNA primer
    4. single-strand binding protein
    Show Answer

    Answer b. Helicase breaks the hydrogen bonds holding together the two strands of DNA.

    Which of these attaches complementary bases to the template strand?

    1. DNA polymerase
    2. helicase
    3. RNA primer
    4. single-strand binding protein
    Show Answer

    Answer a. DNA polymerase builds the new strand of DNA.

    Which of these is later replaced with DNA bases?

    1. DNA polymerase
    2. helicase
    3. RNA primer
    4. single-strand binding protein
    Show Answers

    Answer c. the RNA primer is replaced with DNA nucleotides.

    Learning Objectives

    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.

    Proofreading DNA

    DNA replication is a highly accurate process, but mistakes can occasionally occur, such as a DNA polymerase inserting a wrong base. Uncorrected mistakes may sometimes lead to serious consequences, such as cancer. Repair mechanisms correct the mistakes. In rare cases, mistakes are not corrected, leading to mutations; in other cases, repair enzymes are themselves mutated or defective.

    Most of the mistakes during DNA replication are promptly corrected by DNA polymerase by proofreading the base that has been just added (Figure 7). In proofreading, the DNA pol reads the newly added base before adding the next one, so a correction can be made. The polymerase checks whether the newly added base has paired correctly with the base in the template strand. If it is the right base, the next nucleotide is added. If an incorrect base has been added, the enzyme makes a cut at the phosphodiester bond and releases the wrong nucleotide. This is performed by the exonuclease action of DNA pol III. Once the incorrect nucleotide has been removed, a new one will be added again.

    Illustration shows DNA polymerase replicating a strand of DNA. The enzyme has accidentally inserted G opposite A, resulting in a bulge. The enzyme backs up to fix the error.
    Figure 7. Proofreading by DNA polymerase corrects errors during replication.

    Some errors are not corrected during replication, but are instead corrected after replication is completed; this type of repair is known as mismatch repair (Figure 8). The enzymes recognize the incorrectly added nucleotide and excise it; this is then replaced by the correct base. If this remains uncorrected, it may lead to more permanent damage. How do mismatch repair enzymes recognize which of the two bases is the incorrect one? In E. coli, after replication, the nitrogenous base adenine acquires a methyl group; the parental DNA strand will have methyl groups, whereas the newly synthesized strand lacks them. Thus, DNA polymerase is able to remove the wrongly incorporated bases from the newly synthesized, non-methylated strand. In eukaryotes, the mechanism is not very well understood, but it is believed to involve recognition of unsealed nicks in the new strand, as well as a short-term continuing association of some of the replication proteins with the new daughter strand after replication has completed.

    The top illustration shows a replicated DNA strand with G-T base mismatch. The bottom illustration shows the repaired DNA, which has the correct G-C base pairing.
    Figure 8. In mismatch repair, the incorrectly added base is detected after replication. The mismatch repair proteins detect this base and remove it from the newly synthesized strand by nuclease action. The gap is now filled with the correctly paired base.

    In another type of repair mechanism, nucleotide excision repair, enzymes replace incorrect bases by making a cut on both the 3′ and 5′ ends of the incorrect base (Figure 9).

    Illustration shows a DNA strand in which a thymine dimer has formed. Excision repair enzyme cut out the section of DNA that contains the dimer so it can be replaced with normal base pairs.
    Figure 9. Nucleotide excision repairs thymine dimers. When exposed to UV, thymines lying adjacent to each other can form thymine dimers. In normal cells, they are excised and replaced.

    The segment of DNA is removed and replaced with the correctly paired nucleotides by the action of DNA pol. Once the bases are filled in, the remaining gap is sealed with a phosphodiester linkage catalyzed by DNA ligase. This repair mechanism is often employed when UV exposure causes the formation of pyrimidine dimers.

    Check Your Understanding

    Answer the question(s) below to see how well you understand the topics covered in the previous section. This short quiz does not count toward your grade in the class, and you can retake it an unlimited number of times.

    Use this quiz to check your understanding and decide whether to (1) study the previous section further or (2) move on to the next section.

    https://assessments.lumenlearning.co...sessments/6894

    Contributors and Attributions

    CC licensed content, Original
    • Introduction to DNA Replication. Authored by: Shelli Carter and Lumen Learning. Provided by: Lumen Learning. License: CC BY: Attribution
    CC licensed content, Shared previously
    All rights reserved content
    • DNA replication - 3D. Authored by: yourgenome. Located at: https://youtu.be/TNKWgcFPHqw. License: All Rights Reserved. License Terms: Standard YouTube License

    8.3: DNA Replication is shared under a CC BY-NC-SA license and was authored, remixed, and/or curated by LibreTexts.

    1. Back to top
    • Was this article helpful?

    Recommended articles

    1. Article type
      Section or Page
      License
      CC BY-NC-SA
      OER program or Publisher
      Lumen
    2. Tags
      This page has no tags.