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Eukaryotic DNA Replication – Definition, Steps, Singnification

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Table of Contents

What is Eukaryotic DNA Replication?

  • Eukaryotic DNA replication is a crucial process that ensures the faithful duplication of genetic material in eukaryotic cells. This article provides an in-depth understanding of eukaryotic DNA replication, highlighting its significance and key molecular mechanisms.
  • During DNA replication, DNA polymerases synthesize a complementary DNA strand based on the original template strand. The process begins with DNA helicases unwinding the double-stranded DNA, forming a replication fork that contains two single-stranded templates. This replication fork is where the replication machinery, known as the replisome, coordinates various proteins to carry out the enzymatic functions necessary for DNA synthesis. While the fundamental enzymatic functions are similar in prokaryotes and eukaryotes, eukaryotic DNA replication involves a larger and more complex set of proteins.
  • The replisome is responsible for copying the entire genomic DNA in each dividing cell. This precise replication process ensures the accurate transmission of genetic information from parent cells to daughter cells, making it essential for all organisms. The cell cycle is carefully regulated to ensure error-free DNA replication. In the G1 phase, the regulatory processes for DNA replication initiation are initiated. The majority of DNA synthesis occurs during the S phase, where the entire genome is unwound and duplicated to form two daughter copies. In the G2 phase, any DNA damage or replication errors are corrected. Finally, during mitosis or M phase, one copy of the genomes is segregated into each daughter cell.
  • Eukaryotic DNA replication follows the process of semiconservative replication, which is conserved from prokaryotes. This means that each parental DNA strand acts as a template for the synthesis of a new DNA strand. The replication fork serves as the site for DNA replication, where the DNA helix is unwound, exposing unpaired DNA nucleotides. These nucleotides are recognized and paired with complementary free nucleotides, leading to the formation of double-stranded DNA.
  • It is important to note that eukaryotic DNA replication differs from prokaryotic replication due to the larger size and complexity of eukaryotic genomes. Eukaryotic replication initiation involves multiple initiator proteins, unlike prokaryotic genomes with specific replication sites. However, the overall process remains the same, although different enzymes are involved. The rate of eukaryotic DNA replication is slower compared to prokaryotes, typically around 100 nucleotides per second.
  • In summary, eukaryotic DNA replication is a highly regulated and essential process that ensures the accurate duplication of genetic material. It involves the coordinated actions of various enzymes and proteins to synthesize a complementary DNA strand based on the original template. Understanding the mechanisms of eukaryotic DNA replication is crucial for comprehending how genetic information is inherited and passed on from one generation to the next.

Features of Eukaryotic DNA Replication

Eukaryotic DNA replication exhibits several distinctive features that contribute to its complex nature and accuracy. Understanding these features is essential for comprehending the intricacies of eukaryotic DNA replication. Let’s explore the key characteristics of this process.

  1. Replication Direction: Eukaryotic DNA replication is bi-directional, meaning it occurs in both directions from the origin of replication (Ori C). Multiple origins of replication exist in eukaryotes, facilitating efficient replication throughout the genome.
  2. Semi-Conservative Replication: DNA replication in eukaryotes follows the semi-conservative method. This process results in the formation of a double-stranded DNA molecule consisting of one parental strand and one newly synthesized daughter strand, ensuring the faithful transmission of genetic information.
  3. S Phase and Multiple Origins: Eukaryotic DNA replication exclusively takes place during the S phase of the cell cycle. It occurs at numerous chromosomal origins to efficiently duplicate the entire genome and maintain genomic integrity.
  4. Nuclear Localization: Eukaryotic DNA replication occurs within the cell nucleus. This compartmentalization separates DNA replication from other cellular processes, ensuring efficient and controlled replication.
  5. 5′ to 3′ Synthesis: DNA synthesis during eukaryotic replication exclusively proceeds in the 5′ to 3′ direction. This means that new nucleotides are added to the growing DNA strand in a 5′ to 3′ manner, guided by the template strand.
  6. Leading and Lagging Strands: Eukaryotic DNA replication involves the production of two strands with different synthesis mechanisms. The leading strand is continuously synthesized in the 5′ to 3′ direction, following the replication fork. In contrast, the lagging strand is synthesized in short fragments called Okazaki fragments, which are later joined together.
  7. Polymerases: Eukaryotic cells employ five types of DNA polymerases in the replication process. These polymerases have specific roles and functions, including leading strand synthesis, lagging strand synthesis, DNA repair, and maintaining genomic stability.

What is DNA Polymerases?

DNA polymerases play a crucial role in eukaryotic DNA replication by catalyzing the synthesis of new DNA strands. Eukaryotic cells possess five different DNA polymerases, namely α, β, γ, δ, and ε. Understanding the functions and roles of these polymerases is vital for comprehending the intricacies of eukaryotic DNA replication.

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  1. DNA Polymerases α and δ: DNA polymerases α and δ are primarily involved in replicating chromosomal DNA. Polymerase α carries a primase subunit, allowing it to synthesize RNA primers that initiate the replication process. Polymerase δ then takes over and synthesizes the lagging strand, utilizing the RNA primers as a starting point. This leads to the formation of Okazaki fragments, which are later joined together.
  2. DNA Polymerases β and ε: DNA polymerases β and ε have a role in DNA repair rather than replication. Polymerase β is involved in the repair of damaged DNA, ensuring genomic stability. Polymerase ε also participates in DNA repair processes, contributing to the maintenance of DNA integrity.
  3. DNA Polymerase γ: DNA polymerase γ is unique as it specifically replicates mitochondrial DNA. Mitochondria, which possess their own genetic material, rely on polymerase γ for accurate replication of their DNA.
  4. Telomerase: Telomerase is a specialized DNA polymerase that plays a vital role in replicating DNA at the ends of chromosomes, known as telomeres. Telomerase contains an integral RNA component that serves as its own primer, allowing it to extend and maintain the length of telomeres, which are essential for chromosomal stability.

In addition to DNA polymerases, other enzymes are involved in the DNA replication process:

  1. DNA Topoisomerase I: DNA topoisomerase I relaxes the DNA helix during replication by creating a nick in one of the DNA strands. This relieves the torsional strain on the DNA molecule, facilitating efficient replication.
  2. DNA Topoisomerase II: DNA topoisomerase II relieves the strain on the DNA helix during replication by forming supercoils in the helix. It accomplishes this by creating nicks in both strands of DNA, allowing the DNA molecule to unwind and replicate smoothly.
  3. DNA Ligase: DNA ligase is responsible for sealing the gaps between adjacent DNA fragments during replication. It forms a 3′-5′ phosphodiester bond, joining the fragments together and ensuring the continuity of the DNA strand.

DNA polymerases and associated enzymes work in concert during eukaryotic DNA replication, enabling accurate synthesis, repair, and maintenance of the genetic material. Their coordinated actions contribute to the faithful transmission of genetic information from one generation to the next.

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Eukaryotic DNA replication protein list

ProteinFunction in Eukaryotic DNA Replication
AND1Loads DNA polymerase α onto chromatin together with CMG complex on the lagging strand. Also known as Ctf4 in budding yeast.
Cdc45Required for initiation and elongation steps of DNA replication. A part of the Mcm2-7 helicase complex. Required after pre-RC step for loading of various proteins for initiation and elongation.
Cdc45-Mcm-GINS (CMG) complexFunctional DNA helicase in eukaryotic cells
Cdc6Required for assembly of Mcm2-7 complex at ORC, in conjunction with Cdt1.
Cdc7-Dbf4 kinase or Dbf4-dependent kinase (DDK)Protein kinase required for initiation of DNA replication, probably through phosphorylation of the minichromosome maintenance proteins.
Cdt1Loads Mcm2-7 complex on DNA at ORC in pre-RC/licensing step. Inhibited in metazoans by geminin.
ClaspinCouples leading-strand synthesis with the CMG complex helicase activity. Works with Mrc1.
Ctf4Loads DNA polymerase α onto chromatin together with CMG complex on the lagging strand. Homolog in metazoans is known as AND-1.
Cyclin-dependent kinase (CDK)Cyclin-dependent protein kinase required for initiation of replication and for other subsequent steps.
Dna25′ flap endonuclease and helicase involved in processing Okazaki fragments.
DNA ligase IJoins Okazaki fragments during DNA replication. Ligase activity also needed for DNA repair and recombination.
DNA polymerase α (Pol α)Contains primase activity that is necessary to initiate DNA synthesis on both leading and lagging strands.
DNA polymerase δ (Pol δ)Required to complete synthesis of Okazaki fragments on the lagging strand that have been started by DNA polymerase α.
DNA polymerase ε (Pol ε)The leading strand polymerase. Synthesizes DNA at the replication fork. Binds early at origins via Dbp11 and needed to load DNA polymerase α.
Dpb11DNA replication initiation protein. Loads DNA polymerase ε onto pre-replication complexes at origins.
Fen15′ flap endonuclease involved in processing Okazaki fragments.
GemininProtein found in metazoans and absent from yeasts. Binds to and inactivates Cdt1, thereby regulating pre-replicative/initiation complex formation. Also suggested to promote pre-RC formation by binding and thus preventing Cdt1 degradation.
GINSTetrameric complex composed of Sld5, Psf1, Psf2, Psf3. Associates with pre-replicative complex around the time of initiation and moves with replication forks during the elongation step. Required for the elongation stage of DNA replication and may be part of the Mcm helicase complex.
Minichromosome maintenance proteins (Mcm)Six different proteins of the AAA+ ATPase family that form a hexamer in solution. This hexamer is recruited and loaded by ORC, Cdc6, and Cdt1 and forms a double hexamer that is topologically linked around DNA to form a salt-resistant pre-replicative complex. On replication initiation, Mcm2-7 moves away from ORC with the replication fork.
Mcm10Required for initiation and elongation stages of DNA replication. Implicated in chromatin binding of Cdc45 and DNA polymerase α. Also required for stability of DNA polymerase α catalytic subunit in the budding yeast S. cerevisiae.
Mrc1Couples leading-strand synthesis with the CMG complex helicase activity. Metazoan homolog is known as Claspin.
Origin recognition complex (ORC)Heterohexameric complex composed of Orc1–Orc6 proteins. Binds to DNA and assembles Mcm2-7 complex onto chromatin together with Cdc6 and Cdt1.
Proliferating cell nuclear antigen (PCNA)Trimeric protein with a ring-shaped structure that encloses DNA, preventing dissociation of DNA polymerase. Acts as a sliding clamp for polymerases δ and ε, thereby improving processivity of replicative polymerases.
Replication factor C (RFC)Loads PCNA on primed templates and is involved in the switch between DNA polymerase α and the replicative polymerases δ and ε.
Replication fork barriers (RFBs)Bound by RFB proteins in various locations throughout the genome. Are able to terminate or pause replication forks, stopping the progression of the replisome.
Replication protein A (RPA)Heterotrimeric single-stranded binding protein. Stabilizes single-stranded DNA at the replication fork.
RNase HRibonuclease which digests RNA hybridized to DNA. Involved in Okazaki fragment processing.
Sld2Functions in the initiation of replication. Key substrate of CDK, phosphorylation promotes interaction with Dpb11. Required for initiation of replication.
Sld3Functions in the initiation of replication. Key substrate of CDK, phosphorylation promotes interaction with Dpb11. Required for initiation of replication.
TelomeraseA ribonucleoprotein that adds DNA sequence “TTAGGG” repeats to the 3′ end of DNA strands in telomeres.
TopoisomerasesRegulate the overwinding or underwinding of DNA.

Read Also: Restriction Fragment Length Polymorphism (RFLP)

Initiation of Eukaryotic DNA replication

1. Origin

  • As we saw in the last post, replication starts at a place in the genome called “origin.”
  • The DNA sequences at the replication origin stay the same.
  • Since the eukaryotic genome is a lot bigger than the prokaryotic genome, it has a lot more origins.
  • Because of this, the replication can start at the same time and finish faster.
  • Origin has the ORC’s binding sequence.

The origin recognition complex (ORC)

  • The origin recognition complex (ORC) is a complex of six subunits organised in a C-shaped structure that encircles the DNA at origin.
  • Other elements are bound and recruited into a pre-replicative complex using this complex’s binding and recruitment abilities (pre-RC).

ORC/Cdc6/DNA complex

  • In the later stages of the M phase of the cell cycle, a protein known as cell division cycle 6 (Cdc6) binds the origin of replication (ORC)/origin DNA complex.
  • For the ORC/Cdc6/DNA complex to develop, ATP binding is essential.

OCCM Complex

  • Later, the ORC/Cdc6 complex is bound by the heptamer Cdt1/MCM2-7, which then loads MCM2-7 onto the dsDNA.
  • Minichromosome maintenance 2-7 (MCM2-7) is a fundamental component of the replicative DNA helicase and consists of six subunits stacked in a spiral fashion.
  • Chromatin licencing and DNA replication factor 1, or Cdt1, is responsible for stabilising and loading a Mcm2-7 hexamer onto the chromosome.
  • Cdt1/MCM2-7 binds to the ORC/Cdc6 complex, forming the OCCM complex (fig 1C).

OM intermediate

  • Cdc6 and Cdt1 are released from their cellular encapsulation upon creation of this complex, which also triggers the hydrolysis of ATP.
  • The ORC/MCM2-7 (OM) intermediate is formed when these two proteins are degraded.

OCM complex

  • An additional Cdc6 links to the OM intermediate to form the ORC/Cdc6/MCM2-7 (OCM) complex (fig. 1D), which can then attach a second MCM2-7 hexamer.

DH

  • Cdc6, ORC, and Cdt1 are released while the cell is still in G1 phase as a second MCM2-7 hexamer is recruited to the OCM complex and the two hexamers are aligned to form a head-to-head MCM2-7 double hexamer (DH) (fig 1E).

Pre-replication complex

  • The creation of the pre-replication complex (pre-RC) is a consequence of the DH’s encirclement of the ds DNA.
  • Pre-RC formation might alternatively be thought of as DNA licencing. The RC DH is unable to perform the unwinding actions because it has been dormant since before the RC.

Pre-initiation complex (pre-IC) formation

  • The MCM2-7 DHs are phosphorylated and activated by the kinase DDK during the onset of S phase.
  • Preinitiation complex (pre-IC) creation describes the assembly of proteins with activated MCM2-7.
Eukaryotic initiation of DNA replication
Fig 1: Eukaryotic initiation of DNA replication (Riera et al., 2017)

CMG complex

  • When DH is phosphorylated, a Sld3-binding site on the Mcm2-7 complex becomes accessible. As a result, the Sld3/Sld7 complex and Cdc45 are more likely to attach to replication origins.
  • Sld2 and Sld3 are phosphorylated by cyclin-dependent kinase (CDK) during the S phase of the cell cycle. The phosphoproteins Sld2 and Sld3 interact with the DNA replication regulator protein Dpb11, facilitating its recruitment to the origin of replication.
Fig 2: Unwinding of DNA. (Watase et al., 2012)
Fig 2: Unwinding of DNA. (Watase et al., 2012)
  • Mcm2-7 is bound to accessory factors Cdc45 and GINS via a platform formed by Sld2, Sld3, and Dpb11. Cdc45 and GINS interact to produce the Cdc45/MCM2-7/GINS (CMG) complex, which dissociates Sld2, Sld3, and Dpb11.
Fig 1: Structures of the budding yeast OCCM, DH, and CMG (CMG bound to a replication fork) (Riera et al., 2017).
Fig 1: Structures of the budding yeast OCCM, DH, and CMG (CMG bound to a replication fork) (Riera et al., 2017).
  • Thus, the duplex DNA-encircling MCM2-7 double hexamer undergoes a very controlled transformation into two CMG particles, each of which encircles single-stranded DNA.
  • ATP hydrolysis drives 3′-5′ DNA unwinding, or the helicase activity, and the CMG complex associated with another protein called Mcm10 is the primary component at the eukaryotic DNA replication fork (Fig 1G).
  • Replication protein A (RPA) is a heterotrimeric single-stranded DNA-binding protein that helps stabilise ss DNA as the origin is unwound. Eukaryotic single-stranded DNA binding (SSB) protein is one definition of RPA.
  • Eventually, Pol ε is loaded onto the helicase with the aid of the protein ctf4, following a direct interaction between Pol α and the CMG via GINS. Together, Pol α and DNA primase create a four-subunit complex called DNA Polymerase  α-Primase, which is responsible for RNA primer synthesis and subsequent DNA extension. δ
  • The pol α- primase complex initiates DNA synthesis, and subsequently polymerase δ and ε polymerase take over for the synthesis of the leading and lagging strands, respectively.

Elongation of Eukaryotic DNA replication

  • DNA polymerase α, in the form of DNA polymerase α-primase complex, is the first polymerase to start DNA synthesis.
  • Primers, which are short stretches of RNA between 7 and 12 nucleotides in length, are made by the primase subunit and passed to the polymerase domain, where they are extended with DNA bases (around 20-25 nucleotides).
  • Polymerase switching is a reaction that is started by replication factor C (RFC). DNA polymerase δ and DNA polymerase ε are two more polymerases that help to lengthen the DNA strands.

Leading strand synthesis

  • After DNA pol α has finished synthesising the RNA primer and adding the DNA bases, the RFC dissociates DNA pol α and triggers the assembly of proliferating cell nuclear antigen (PCNA) at the primer terminus region. DNA polymerases use PCNA as a DNA clamp.
Leading strand synthesis
Leading strand synthesis
  • It has been found that Pol  ε  is the primary polymerase involved in leading strand synthesis (in Saccharomyces cerevisiae). Since the extension occurs in the leading strand and replication is continuous, a single round of primer synthesis is sufficient (fig 1).

Lagging strand synthesis

Lagging strand synthesis
Lagging strand synthesis
  • Polymerases δ is the main polymerase in lagging-strand synthesis. The replication in the lagging strands is discontinuous and takes place with production of numerous Okazaki fragments (fig 2).
Elongation
Okazaki fragments
  • The Okazaki fragments (fig. 3) produced by lagging-strand synthesis in eukaryotes are on the order of 200 bases in length, but in bacteria they are on the order of 2000 bases. Because of the increasing number of Okazaki fragments synthesised, polymerase switching during Okazaki fragment synthesis has taken on more significance.
  • Therefore, DNA pol α adds about 10–20 nucleotides of DNA bases to an initial RNA (7–10 nucleotides) in an Okazaki fragment.
  • Deoxyribonucleotides are incorporated into strands of DNA by DNA pol δ while the polymerase switching triggered by the RFC is maintained in place by the sliding clamp PCNA (see the figure below). Upon reaching the preceding RNA primer, DNA pol δ dissociates, indicating that DNA synthesis is complete.
  • By degrading RNA primers, RNase H1 leaves only one ribonucleotide bound to the DNA (3′ end) of the Okazaki fragment. Flap endonuclease 1 then eliminates the ribonucleotide that was overlooked (FEN 1).
The proteins involved in the replication, especially PCNA (Boehm et al., 2016).
The proteins involved in the replication, especially PCNA (Boehm et al., 2016).
  • After the primers are removed, Okazaki fragments are left with gaps in between them, which are filled by polymerase. δ DNA ligase I bridges the nick between the Okazaki fragment and the lagging strand, creating a continuous lagging strand.

Nucleosome Assembly

  • The nucleosome packing along the DNA undergoes continual disintegration and reassembly during elongation. One characteristic of eukaryotic DNA is that it is bundled into extremely tiny structures called chromosomes.
  • This process involves nucleosome formation, in which negatively charged DNA is wound around basic proteins called histones (fig below).
  • There are 8 histone proteins in a nucleosome, 2 each of H2A, H2B, H3, and H4. Between two nucleosomes, histone H1 acts as a linker.
  • DNA replication destabilises the two nucleosomes that sit on the strand of unreplicated DNA immediately in front of the replication fork.
  • Therefore, DNA packaging becomes disorganised as a result of replication fork movement, allowing replication proteins to interact with the DNA.
  • In the duplicated region, nucleosome-free regions extended for around 225 base pairs (bp) and 285 bp (bp) on the leading and lagging strands, respectively. Histone octomer encased the DNA after this area, however part of the chromosomes were missing histone H1.
  • Complete nucleosomes containing H1 were found in the daughter strands after approximately 450 bp to 650 bp had passed from the replication fork.
  • Therefore, the nucleosome is constructed by the sequential assembly of daughter strands.
  • In this process, the parental nucleosomes separate and bind to the daughter DNA strands at random, creating a situation in which each strand receives one-half of the nucleosomes from the other.
  • When the two daughter strands are ready for packing, the remaining nucleosome components are generated from scratch and placed onto the strands.
Nucleosome Assembly
Nucleosome Assembly | David O Morgan, Attribution, via Wikimedia Commons

Termination of Eukaryotic DNA Replication

If you want two distinct copies of your DNA, replication must be stopped after it has been completed successfully.

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1. Involving two adjacent replication forks

Termination in eukaryotic cells requires the merger of two neighbouring replication forks due to the high number of origins present in these cells. The process entails the following four stages:

Dissolution

  • Both CMGs that are moving toward each other unwind the DNA segment between the two forks in Fig. 5.a (Fig. 5.b) (fig 5.c). DNA pol δ and the elongation factor FEN1 work together to process the final Okazaki fragment (fig 5.d).

Ligation

  • Two synthesised strands, one approaching from the opposite direction, are ligated together, closing the gaps in the daughter strands (as with regular Okazaki fragments).

Dissociation of replisome

  • Following the merger of the two replication forks, the replisome complex is disassembled. Mcm7 and the p97/VCP/Cdc48 segregase are polyubiquitylated at the end of the process (fig 5.e).

Decatenation

  • Topoisomerase II is used to separate the two strands of DNA if there are any entanglements, also known as catenanes, in the daughter strands.
Fig 5: Termination involves merging of the two neighbouring forks (Dewar & Walter, 2017).
Fig 5: Termination involves merging of the two neighbouring forks (Dewar & Walter, 2017).

2. Involving the ends of the Chromosomes

Fig 6: Telomeres form protective end of eukaryotic linear DNA (Aulinas, 2013).
Fig 6: Telomeres form protective end of eukaryotic linear DNA (Aulinas, 2013).
  • It is well known that the DNA in eukaryotic chromosomes is a linear molecule, and that the process of terminating eukaryotic DNA involves completing replication at the ends of chromosomes, which are called Telomeres (fig 6).
  • The 3′-OH group from an RNA primer facilitates 5′-to-3′ replication during the creation of Okazaki fragments. After the RNA primer is removed from the unreplicated chromosomal end, the length of the freshly synthesised strand is reduced (fig 7).
Shortening of the chromosome ends.
Shortening of the chromosome ends.
  • Telomeres (and telomere-associated proteins) are repeating sequences found at the ends of chromosome DNA that protect them from being shortened. Such as a human, At birth, chromosomes have telomeres that are 15-20 kb long and include repeating sequences of (TTAGGG)n. Chromosome ends are safeguarded by these structures to prevent false positives for DNA double strand breaks at their location (DSB).
  • When DNA replicates normally in somatic cells, the telomeric region of eukaryotic chromosomes shortens. Replicative senescence or apoptosis occurs when telomeres get too short after a specific number of DNA replications and, thus, cell divisions.
  • However, an enzyme called telomerase stretches chromosomal ends, particularly the 5′ end of lagging strands, in germline and cancer cells. These cells are immortal because they are able to keep their telomeres at a constant length.
  • Template-encoding RNA molecule (TER) refers to the RNA template used by the reverse transcriptase telomerase (fig 8). TERT is the short form of telomerase RNA-binding protein (TElomerase Reverse Transcriptase).
  • Our human RNA template consists of the letters AUCCCAAUC. Telomeric DNA repeats are produced and appended to the 3′ overhang of single-stranded DNA (fig 8).
  • DNA pol and DNA ligase finish DNA strand synthesis once telomerase has finished extending the 3′ end of the strand.
Fig 8: Synthesis of telomeric DNA repeats by Telomerase (Verhoeven et al, 2014).
Fig 8: Synthesis of telomeric DNA repeats by Telomerase (Verhoeven et al, 2014).

DNA Proofreading

  • DNA Proofreading is a crucial mechanism that ensures the accuracy of DNA replication and maintenance of genetic information. In eukaryotes, specific polymerases involved in DNA elongation, such as delta and epsilon polymerases, possess a built-in proofreading ability through their 3′ to 5′ exonuclease activity.
  • During DNA replication, these polymerases move along the template strand, adding complementary nucleotides to the growing daughter strand. However, occasionally, errors can occur, resulting in the insertion of incorrect nucleotides. To prevent the perpetuation of these errors, the proofreading function of the polymerases comes into play.
  • When an error is detected during DNA replication, the polymerase pauses its activity. The 3′ to 5′ exonuclease activity of the polymerase allows it to backtrack along the newly synthesized strand. This exonuclease activity enables the polymerase to remove the incorrect nucleotide, effectively excising it from the growing strand.
  • Once the erroneous nucleotide is removed, the polymerase resumes its elongation activity, adding the correct nucleotide in its place. This process ensures that the DNA sequence is accurately replicated, maintaining the integrity of the genetic information.
  • In addition to proofreading during replication, cells have mechanisms to repair DNA damage caused by various sources, including environmental factors like UV radiation. One of the repair mechanisms employed is excision repair.
  • Excision repair is responsible for removing and replacing specific types of DNA lesions, such as pyrimidine dimers induced by UV rays or other mutated bases. This repair process involves recognition and excision of the damaged DNA segment, followed by the synthesis of a new strand with the correct sequence. Excision repair helps to maintain the integrity of the DNA by fixing damaged regions and preserving the accurate genetic code.
  • In summary, DNA proofreading is a vital mechanism that corrects errors made during DNA replication, ensuring the fidelity of genetic information. The 3′ to 5′ exonuclease activity of specific polymerases allows for the removal of incorrect nucleotides and their replacement with the correct ones. Furthermore, excision repair is a repair mechanism that removes and replaces damaged DNA segments, safeguarding the integrity of the genetic material.

Differences Between Prokaryotic DNA replication and Eukaryotic DNA replication

Prokaryotic DNA replicationEukaryotic DNA replication
Occurs inside the cytoplasmOccurs inside the nucleus
Only one origin of replication per molecule of DNAHave many origins of replication in each chromosome
Origin of replication is about 100-200 or more nucleotides in lengthEach origin of replication is formed of about 150 nucleotides
Replication occurs at one point in each chromosomeReplication occurs at several points simultaneously in each chromosome
Only have one origin of replicationHas multiple origins of replication
Initiation is carried out by protein DnaA and DnaBInitiation is carried out by the Origin Recognition Complex
Topoisomerase is neededTopoisomerase is needed
Replication is very rapidReplication is very slow

Significance of Eukaryotic DNA Replication

  • The significance of eukaryotic DNA replication lies in its essential role in cell growth, division, and the conservation of genetic information. DNA replication is a fundamental process that generates new molecules of DNA, which are crucial for the maintenance and transmission of genetic material.
  • One of the primary functions of DNA replication is to ensure the accurate transmission of genetic information from one generation to the next. Each eukaryotic cell contains a complete set of chromosomes comprising the entire genome. Through DNA replication, an identical copy of the genome is produced, ensuring that each daughter cell receives a complete and accurate set of genetic instructions.
  • Proper regulation of DNA replication is vital for the growth and division of cells. During cell division, DNA replication occurs to ensure that each daughter cell receives the necessary genetic material. The precise timing and coordination of DNA replication with other cellular processes are crucial for maintaining the integrity of the genome and facilitating successful cell division.
  • Moreover, DNA replication plays a significant role in various cellular processes, including DNA repair, recombination, and gene expression. Replication provides an opportunity for DNA damage to be detected and repaired, ensuring the stability of the genome. It also enables the exchange of genetic material between homologous chromosomes during recombination, contributing to genetic diversity.
  • In addition to its role in cell growth and division, DNA replication is essential for the development and differentiation of multicellular organisms. During embryonic development, DNA replication is intricately regulated to support the generation of specialized cell types with specific functions. This process ensures that each cell type possesses the appropriate genetic information necessary for its unique role within the organism.
  • In summary, the significance of eukaryotic DNA replication lies in its crucial role in cell growth, division, and the conservation of genetic information. By accurately replicating the entire genome, DNA replication ensures the transmission of genetic material to subsequent generations. It also contributes to DNA repair, recombination, and gene expression, and supports the development and differentiation of multicellular organisms. Understanding the intricacies of DNA replication is essential for unraveling the mechanisms underlying cellular processes and genetic inheritance.

FAQ

What is eukaryotic DNA replication?

Eukaryotic DNA replication is the process by which a eukaryotic cell duplicates its genetic material, specifically the DNA molecules that make up the chromosomes.

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How does eukaryotic DNA replication differ from prokaryotic DNA replication?

Unlike prokaryotes, eukaryotic DNA replication occurs within the nucleus of the cell rather than in the cytoplasm. Eukaryotes also have multiple origins of replication per chromosome, while prokaryotes typically have only one.

What is the purpose of eukaryotic DNA replication?

The primary purpose of eukaryotic DNA replication is to ensure the accurate transmission of genetic information from one generation of cells to the next. It is essential for cell growth, division, and the maintenance of genetic integrity.

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Which enzymes are involved in eukaryotic DNA replication?

Several enzymes play key roles in eukaryotic DNA replication, including DNA polymerases, helicases, topoisomerases, and ligases. These enzymes work together to unwind the DNA, synthesize new strands, and ensure proper DNA strand rejoining.

How does eukaryotic DNA replication ensure accuracy?

Eukaryotic DNA replication incorporates proofreading mechanisms to ensure accuracy. The DNA polymerases involved in replication possess exonuclease activity, allowing them to detect and remove incorrect nucleotides before continuing with replication.

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What is the role of origins of replication in eukaryotic DNA replication?

Origins of replication are specific sites on the DNA where replication initiates. In eukaryotes, multiple origins of replication are present on each chromosome to facilitate efficient and timely replication.

How does eukaryotic DNA replication coordinate with other cellular processes?

Eukaryotic DNA replication is tightly regulated and coordinated with other cellular processes, such as cell cycle progression and DNA repair. This ensures that DNA replication occurs at the appropriate time and is accurately completed.

Are there any factors that can affect eukaryotic DNA replication?

Yes, various factors can influence eukaryotic DNA replication. Environmental factors, DNA damage, and mutations in replication-related genes can all impact the efficiency and fidelity of DNA replication.

Can errors occur during eukaryotic DNA replication?

Although eukaryotic DNA replication is highly accurate, errors can still occur. These errors can result in mutations that may have consequences for cellular function and potentially lead to genetic disorders or diseases.

How is eukaryotic DNA replication related to cancer?

Defects in DNA replication and repair mechanisms can contribute to the development of cancer. Mutations in genes involved in replication and checkpoint control can disrupt the normal regulation of cell growth, leading to uncontrolled cell division and tumor formation.

References

  • Bhagavan, N. V., & Ha, C.-E. (2015). DNA Replication, Repair, and Mutagenesis. Essentials of Medical Biochemistry, 401–417. doi:10.1016/b978-0-12-416687-5.00022-1 
  • Priego, Sara & Gambus, Aga. (2015). Regulation of Unperturbed DNA Replication by Ubiquitylation. Genes. 6. 451-68. 10.3390/genes6030451. 
  • Riera et al. (2017) From structure to mechanism- understanding initiation of DNA replication. Genes Dev. 31(11): 1073-1088. doi: 10.1101/gad.298232.117
  • Bhagavan & Ha (2015) Chapter 22 – DNA Replication, Repair, and Mutagenesis. Essentials of Medical Biochemistry (Second Edition) With Clinical Cases. 401-417. https://doi.org/10.1016/B978-0-12-416687-5.00022-1
  • Tanaka et al. (2011) Sld7, an Sld3-associated protein required for efficient chromosomal DNA replication in budding yeast. EMBO J. 30(10): 2019-30. doi: 10.1038/emboj.2011.115. Epub 2011 Apr 12.
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