Name: dna replication teams in the replisome
Uploaded: 2020-11-23T11:47:35.000Z
Duration: 3 min 1 s
Channel: JoVE

K12 Standards

Alabama Course of Study Science

9-12

Biology

from-molecules-to-organisms-structures-and-processes

From Molecules to Organisms: Structures and Processes

9-12.B.1.3

The Replisome

A replicação do DNA é realizada por uma montagem multiproteina altamente coordenada, conhecida como a máquina de replicação do DNA, ou o replissomo, o que aumenta a eficiência da replicação do DNA. Os componentes do núcleo da maquinaria são helicases, proteínas de ligação do DNA de uma só cadeia, DNA primasse, grampos deslizantes, um carregador de fechamento, e DNA múltiplo DNA primasse, que são todos associados uns aos outros próximo ao garfo de replicação. As polimerases do DNA Replicativo têm um processamento de ao redor de 10 nucleotídeos, que é o número de nucleotídeos que se pode adicionar à vertente da filha antes de dissociar 00:00:45.070 00:00 48.160 do modelo.Isto é demasiado ineficiente para copiar genomas inteiros em um período de tempo razoável, e este problema é resolvido com a ajuda de proteínas de fechamento deslizantes. Quando ATP associa-se com a proteína do carregador do grampo, ligarão e abrirão a braçadeira deslizante, de modo que sua estrutura em forma de anel pode circundar o complexo do primer-DNA do molde. Uma vez ligado, o grampo de fechamento hidrolisa o ATP à ADP, fazendo com que o grampo de fechamento desassocie e o grampo para fechar próximo ao DNA.Então, a DNA polimerase se liga às proteínas de fechamento. E juntas, deslizam ao longo do DNA modelo, a polimerase do DNA das amarras à fita e aumentando seu processamento em até 1, 000 nucleotídeos. Este aumento do processamento permite que a DNA polimerase realize a replicação contínua do DNA na vertente na principal.No entanto, no modelo de vertente mais atrasada, outra polimerase do DNA executa a replicação descontínua do DNA de uma maneira que permite que as moléculas de DNA polimerase sintetizem em simultâneo as cadeias principais e atrasadas. Este processo é descrito às vezes como o modelo do trombone. A vertente mais atrasada e seu molde de tira formam um laço quando DNA polimerase inicia a síntese do fragmento de Okazaki de um primer do RNA.A fita DNA cresce de ambas as direções quando o DNA se desdobra da helicase e a vertente mais atrasada é sintetizada. Quando a DNA polimerase encontra o próximo RNA primer, se separa da cadeia modelo. Entretanto, a primasse adiciona um outro primer à vertente mais atrasada.E a fita crescente do DNA é liberada. As proteínas do grampo de fechamento e grampo permitem DNA polimerase rapidamente associe com o modelo de DNA preparado. A formação e posterior colapso da fita de DNA se repete com a síntese de cada novo fragmento de Okazaki.

<ol>
	<li>O'Donnell, Michael, Lance Langston e Bruce Stillman. "Princípios e conceitos de replicação de DNA em bactérias, archaea e eukarya." Cold Spring Harbor Perspectives in Biology 5, no. 7 (2013): a010108.</li>
	<li>"Replicação e recombinação de DNA." Natureza 421, nº 6921 (2003): 431.</li>
	<li>Kelch, Brian A., Debora L. Makino, Mike O'Donnell e John Kuriyan. "ATPases de carregador de grampo e a evolução da maquinaria de replicação de DNA." BMC Biology 10, nº 1 (2012): 34.</li>
	<li>Hedglin, Mark, Ravindra Kumar e Stephen J. Benkovic. "Grampos de replicação e carregadores de grampos." Perspectivas de Cold Spring Harbor em Biologia 5, nº 4 (2013): a010165.</li>
	<li>Gardner, Andrew F. e Zvi Kelman. "A maquinaria de replicação do DNA como alvos terapêuticos." Fronteiras em Biociências Moleculares 6 (2019): 35.</li>
</ol>

o replísomo

A replicação do DNA é realizada por um grande complexo de proteínas que atuam de forma coordenada para alcançar a replicação do DNA de alta fidelidade. Juntos, esse complexo é conhecido como maquinaria de replicação do DNA ou replissoma. A síntese das vertentes principal e atrasada é um processo altamente coordenado. Para explicar isso, o “modelo Trombone” foi proposto por Bruce Alberts em 1980. A formação da alça de DNA começa quando um primer é sintetizado na fita parental atrasada. O loop cresce com a síntese de um fragmento de Okazaki e finalmente se dissolve quando a síntese do fragmento termina. A DNA polimerase então se liga a outro primer e todo o ciclo, desde a formação da alça até o colapso, é repetido. O replissoma permite que todas as atividades sejam coordenadas como um complexo de máquinas de replicação. Componentes responsivos As helicases se desenrolam e separam o DNA de fita dupla. Essas enzimas estão presentes como um anel hexâmero único em procariontes e como um anel hexâmero duplo em eucariotos. As helicases eucarióticas dependem de proteínas adicionais, Cdc45 e GINS, para funcionar. As proteínas de ligação ao DNA de fita simples (SSB) evitam que as fitas de DNA se recombinem. Nos procariontes, os SSBs consistem em uma única subunidade, enquanto nos eucariotos são uma proteína heterotrimérica conhecida como proteína de replicação A (RPA). Primase adiciona um primer de RNA ao local onde a síntese de DNA se originará. Nos procariontes, a primase está presente como uma enzima de subunidade única chamada DnaG, que sintetiza um primer de RNA de cerca de 12 nucleotídeos. Em eucariotos, uma enzima multisubunidade, DNA polimerase-α primase, sintetiza um primer híbrido RNA-DNA de cerca de 25 nucleotídeos. Além da polimerase-α, a polimerase replicativa estenderá o DNA recém-sintetizado. Embora um único tipo de polimerase replicativa, a DNA polimerase III, esteja presente em procariontes, dois tipos diferentes de polimerases replicativas, Pol ε e Pol δ, estão presentes em eucariotos, para síntese de fita principal e de fita retardada, respectivamente. As proteínas Sliding Clamp mantêm as polimerases ligadas ao modelo de DNA. β-clamp, uma proteína homodimérica, atua como clamp em procariontes. Nos eucariotos, a mesma tarefa é realizada pelo antígeno nuclear celular em proliferação (PCNA), uma proteína homotrimérica. O poro central da pinça deslizante é carregado positivamente, o que lhe permite ter interações eletrostáticas com a estrutura de fosfato carregada negativamente do DNA. O grampo é anexado ao DNA por um Clamp Loader. Essas proteínas pentaméricas pertencem à classe AAA+ de ATPases. O clamp loader eucariótico é conhecido como fator de replicação C, enquanto o clamp loader procarionte de E. coli é conhecido como complexo γ; no entanto, acredita-se que as proteínas clamp e os carregadores clamp sejam homólogos evolutivos, ao contrário de muitos outros componentes do replissoma.

O Replísomo

A replicação do DNA é realizada por um grande complexo de proteínas que atuam de forma coordenada para alcançar a replicação do DNA de alta fidelidade. Juntos, isso ...

Education

JoVE Core

Molecular Biology

Biologia Molecular

Unit 1

Unidade 1

DNA Replication

Replicação do DNA

KEY TERMS AND CONCEPTS

TERMOS-CHAVE E CONCEITOS

dna replication teams in the replisome

DNA replication depends on the replisome, a coordinated complex of proteins that copies DNA with high accuracy. This DNA replication machinery brings many parts together so the leading and lagging strands can be made at the same time.

A key idea in this process is the trombone model, proposed by Bruce Alberts in 1980. In this model, a loop forms on the lagging strand when a primer is made. The loop grows as an Okazaki fragment is synthesized, then falls apart when that fragment is finished. DNA polymerase then moves to a new primer, and the cycle repeats.

Helicases start the process by unwinding and separating double-stranded DNA. In prokaryotes, helicase is a single hexamer ring, while in eukaryotes it forms a double hexamer ring. Eukaryotic helicases also need the extra proteins Cdc45 and GINS to work. Single-stranded DNA binding proteins keep the separated strands from reannealing. In prokaryotes, these are SSB proteins with one subunit. In eukaryotes, the equivalent protein is replication protein A, or RPA, which is a heterotrimer.

Primase adds the RNA primer that gives DNA synthesis a starting point. In prokaryotes, primase is the single-subunit enzyme DnaG, and it makes an RNA primer of about 12 nucleotides. In eukaryotes, DNA polymerase-alpha primase makes an RNA-DNA hybrid primer of about 25 nucleotides. After that, the replicative polymerase extends the new DNA strand. Prokaryotes use DNA polymerase III, while eukaryotes use Pol epsilon for the leading strand and Pol delta for the lagging strand.

Sliding clamp proteins hold the polymerases on the DNA template so copying can continue efficiently. The prokaryotic beta-clamp is a homodimer, and the eukaryotic clamp is proliferating cell nuclear antigen, or PCNA, which is a homotrimer. The clamp has a positively charged central pore that interacts with the negatively charged DNA backbone. A clamp loader places the clamp onto DNA. These pentameric AAA+ ATPases include the eukaryotic replication factor C and the E. coli gamma complex. The clamp proteins and clamp loaders are thought to be evolutionary homologs, unlike many other replisome components.

DNA Replication Teams in the Replisome

replication in prokaryotes

Prokaryotic DNA Replication Steps

Prokaryotic DNA replication copies the chromosome before a cell divides. In bacteria, this process starts at a specific DNA site called the origin of replication. From there, the cell builds new DNA strands using enzymes that match each base with its partner. The DNA double helix first opens so replication can begin. As the strands separate, each original strand serves as a template. New nucleotides are added in a set order, which helps the cell make an accurate copy of its genetic material. <br...

replication in eukaryotes

Eukaryotic DNA Replication Steps

Eukaryotic DNA replication copies the genome before a cell divides. In eukaryotes, this process happens in a controlled sequence so each new cell can receive genetic information. Replication begins at specific sites on the DNA called origins. The DNA is opened, and each original strand serves as a template, meaning it guides the building of a new complementary strand. This sets up two DNA molecules from one original molecule. The process then continues as enzymes add matching nucleotides to...

dna base pairing

DNA Base Pair Rules

DNA base pairing follows specific rules that help DNA carry genetic information. In DNA, the bases pair in a regular way, with adenine matching thymine and cytosine matching guanine. These pairs fit together because of their chemical shapes and bonding patterns. These base pairs also support accurate DNA replication. When DNA copies itself, each strand can guide the formation of a new partner strand using the pairing rules. Understanding base pairing helps explain how DNA stays organized and...

the dna replication fork

DNA Replication Stress and Fork Stalling

DNA replication stress and fork stalling can disrupt the copying of a cell’s genome. The replication fork is the Y-shaped site where DNA strands separate and new complementary strands are made at the same time. This linked unwinding and synthesis helps cells duplicate DNA efficiently and with fewer errors. In organisms with small circular DNA, such as E. coli, replication often begins at a single origin. That setup creates two replication forks that move in opposite directions from the opening...

proofreading

Proofreading in DNA Replication

Proofreading in DNA replication helps keep DNA copying accurate. It is a built-in check that reduces mistakes as new DNA is made. DNA polymerase is the enzyme that carries out this checking during replication. When it adds a wrong nucleotide, it can remove the error and replace it with the correct one. This correction step helps maintain the correct DNA sequence. Proofreading is an important part of DNA replication because it supports genetic accuracy. By catching mistakes as they happen, the...

dna helicases

Helicase Motor Proteins in DNA Unwinding

Helicase motor proteins help unwind DNA during key cell processes. They use energy from ATP hydrolysis, which is the breakdown of ATP to release usable energy. This movement lets helicases separate DNA strands when the cell needs access to the genetic code. These enzymes are involved in DNA replication, repair, recombination, and transcription. Helicases are found in all living organisms, but their structure, function, and movement can differ. In prokaryotes, the DnaB helicase moves along the...

lagging strand synthesis

Okazaki Fragments in DNA Replication

DNA replication uses a leading strand and a lagging strand, and they are made at different rates. The leading strand starts first. The lagging strand starts later, moves more slowly, and is made in pieces. The two strands also differ in direction and starting steps. Leading strand synthesis follows the opening replication fork. Lagging strand synthesis goes in the opposite direction. The leading strand needs one RNA primer, while the lagging strand needs multiple RNA primers. After priming,...

the replisome

DNA replication depends on the replisome, a coordinated complex of proteins that copies DNA with high accuracy. This DNA replication machinery brings many parts together so the leading and lagging strands can be made at the same time. A key idea in this process is the trombone model, proposed by Bruce Alberts in 1980. In this model, a loop forms on the lagging strand when a primer is made. The loop grows as an Okazaki fragment is synthesized, then falls apart when that fragment is finished.

mismatch repair

Fixing DNA Copying Errors

Mismatch repair helps fix DNA copying errors after replication. When DNA is copied, small mistakes can slip through. This repair system finds those mismatches and corrects them before they become permanent changes. The process depends on a group of mutator proteins. These proteins work together to detect the error, remove the incorrect section, and help replace it with the proper DNA sequence. In this way, mismatch repair supports the accuracy of genetic information. If the repair system does...

dna topoisomerases

How Topoisomerases Untwist DNA

DNA topoisomerases are enzymes that untwist DNA when it becomes overcoiled during cell processes such as DNA replication and transcription. They help regulate positive and negative DNA supercoiling without changing the nucleotide sequence. Clockwise overwinding creates positively supercoiled DNA, while counterclockwise underwinding produces negatively supercoiled DNA. Topoisomerases are grouped into two main types. Type I topoisomerases act on one strand of double-stranded DNA, and they...

telomeres and telomerase

How Telomerase Protects Chromosome Ends

Telomerase helps protect chromosome ends during eukaryotic DNA replication. When the final primer is removed, a single-stranded DNA fragment can remain at the end of the chromosome. That piece cannot be copied in the same way as the rest of the strand because there is no 3' end for new DNA to attach to. If this end piece is not replaced, chromosomal DNA is gradually lost each time a cell divides. The exposed single-stranded DNA can also trigger a DNA damage response from enzymes that detect it.

non-nuclear inheritance

Cytoplasmic Inheritance in Cells

Cytoplasmic inheritance explains how some traits are passed through cell structures outside the nucleus. These traits come from organelles such as mitochondria and chloroplasts. They do not follow the usual nuclear pattern of inheritance. Mitochondria and chloroplasts each contain their own DNA. Because of this, they can carry genetic information that affects cell function and certain visible traits. In high school biology, this topic helps explain why not all inherited traits come from...

animal mitochondrial genetics

Animal Mitochondrial DNA and Inheritance

Animal mitochondrial DNA has its own genome, separate from nuclear DNA. It is a double-stranded, closed circular molecule with about 20,000 base pairs. One strand is the heavy, or H-strand, because it is rich in guanine. The other is the light, or L-strand, because it is rich in cytosine. Compared with nuclear DNA, mitochondrial DNA has very little non-coding sequence. It also has no introns. Its genes are packed closely together, and some even overlap. The D-loop is the main non-coding...

comparing mitochondrial, chloroplast, and prokaryotic genomes

comparing-mitochondrial-chloroplast-and-prokaryotic-genomes

How Organelle Genomes Resemble Bacteria

Mitochondrial and chloroplast genomes still keep several traits from their bacterial ancestors. They also show changes that happened after these organelles evolved inside eukaryotic cells. Eukaryotic cells are cells with a nucleus, while prokaryotes are cells without one. Like prokaryotic genomes, mitochondrial and chloroplast genomes do not bind with histone-like proteins. They also do not show the complex chromosome-like packaging seen in eukaryotes. Mitochondria and chloroplasts divide by...

export of mitochondrial and chloroplast genes

export-of-mitochondrial-and-chloroplast-genes

How Organelles Pass Genes to the Nucleus

Eukaryotic cells can carry three genetic systems: nuclear, mitochondrial, and chloroplast. Over time, many genes from mitochondria and chloroplasts have moved into the nucleus. In some plant species, this gene transfer is still happening. This movement has affected large parts of the genome in some organisms. About 18% of the Arabidopsis thaliana nuclear genome is thought to come from the chloroplast's cyanobacterial ancestor. In yeast, around 75% of the genome is derived from the...

single-strand dna binding proteins

How SSB Proteins Protect DNA Strands

Single-strand DNA-binding proteins, or SSB proteins, help protect DNA during replication. When double-stranded DNA is unwound, the two strands separate. SSB proteins bind to the exposed single strands and keep them stable. These proteins bind in a sequence-independent manner. That means the order of the nitrogenous bases does not have to follow a specific pattern for SSB proteins to attach. Their binding straightens single-stranded DNA and makes it more rigid. This support may help DNA...

key terms and concepts

Explore how the replisome coordinates DNA replication, from helicase and primase to polymerases, sliding clamps, and the trombone model.

DNA Replication Teams in the Replisome | JoVE K12

molecular biology

JoVE Core Biologia Molecular mostra os processos moleculares da célula através de vídeos de animação claros e concisos. Além disso, nos vídeos de Cientistas em ação, a aplicação desses conceitos é demonstrada por meio de experimentos reais em laboratórios modernos.

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9-12.B.1.1

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Transcription

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Eukaryotic RNA Polymerases

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RNA Polymerase II Accessory Proteins

Transcription Elongation Factors

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Pre-mRNA Processing: RNA Splicing

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Alternative RNA Splicing

Nuclear Export of mRNA

Transfer RNA Synthesis

Ribosomal RNA Synthesis

The Nucleolus

Additional Subnuclear Structures

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tRNA Activation

Initiation of Translation

Termination of Translation

Improving Translational Accuracy

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Nonsense-mediated mRNA Decay

The Eukaryotic Promoter Region

Molecular Chaperones and Protein Folding

Co-activators and Co-repressors

The Proteasome

Master Transcription Regulators

Regulated Protein Degradation

Regulated mRNA Transport

Proteins: From Genes to Degradation

mRNA Stability and Gene Expression

Cell Specific Gene Expression

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Cis-regulatory Sequences

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9-12.B.1.2

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a. Use a model to describe the structure and sequence of DNA, including nucleotide structure, base pairing, and the structure of the helix.

9-12.B.1.2.a

DNA Replication Teams in the Replisome

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