Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Ziegler–Natta Chain-Growth Polymerization: Overview01:17

Ziegler–Natta Chain-Growth Polymerization: Overview

3.4K
Ziegler–Natta polymerization is another form of addition or chain‐growth polymerization used for synthesizing linear polymers over branched polymers. The catalyst used for polymerization is the Ziegler–Natta catalyst, named after Karl Ziegler and Giulio Natta, who developed it in 1953. This catalyst is an organometallic complex of titanium tetrachloride and triethyl aluminum, with the active form of the catalyst being an alkyl titanium compound. Using the Ziegler–Natta...
3.4K
Bacterial Protein Maturation01:26

Bacterial Protein Maturation

36
Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
36
Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

2.3K
2.3K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

5.8K
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
5.8K
Lagging Strand Synthesis01:59

Lagging Strand Synthesis

53.0K
During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
53.0K
Anionic Chain-Growth Polymerization: Mechanism01:04

Anionic Chain-Growth Polymerization: Mechanism

2.1K
The mechanism for anionic chain-growth polymerization involves initiation, propagation, and termination steps. In the initiation step, a nucleophilic anion, such as butyl lithium, initiates the polymerization process by attacking the π bond of the vinylic monomer. As a result, a carbanion, stabilized by the electron‐withdrawing group, is generated. The resulting carbanion acts as a Michael donor in the propagation step and attacks the second vinylic monomer, which acts as a Michael...
2.1K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Identification of determinants of high-fidelity DNA synthesis in Mycobacterium smegmatis DnaE1 through in silico and in vivo approaches.

Nucleic acids research·2025
Same author

A unique inhibitor conformation selectively targets the DNA polymerase PolC of Gram-positive priority pathogens.

Nature communications·2025
Same author

DNA polymerase actively and sequentially displaces single-stranded DNA-binding proteins.

Nature communications·2025
Same author

Structural and functional analysis of the <i>Mycobacterium tuberculosis</i> MmpS5L5 efflux pump presages a pathway to increased bedaquiline resistance.

bioRxiv : the preprint server for biology·2025
Same author

NAD-dependent dehydrogenases enable efficient growth of Paracoccus denitrificans on the PET monomer ethylene glycol.

Nature communications·2025
Same author

Investigating the composition and recruitment of the mycobacterial ImuA'-ImuB-DnaE2 mutasome.

eLife·2023

Related Experiment Video

Updated: Jul 18, 2025

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

1.9K

A four-point molecular handover during Okazaki maturation.

Margherita M Botto1,2, Alessandro Borsellini1,3, Meindert H Lamers4

  • 1Department of Cell and Chemical Biology, Leiden University Medical Center (LUMC), Leiden, the Netherlands.

Nature Structural & Molecular Biology
|August 24, 2023
PubMed
Summary

DNA replication requires precise coordination of enzymes to remove RNA primers from the lagging strand. This study reveals a four-step molecular handover mechanism ensuring efficient primer removal and DNA synthesis completion in E. coli.

More Related Videos

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
06:24

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Published on: February 13, 2019

8.1K
Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
07:27

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

13.6K

Related Experiment Videos

Last Updated: Jul 18, 2025

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

1.9K
Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
06:24

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Published on: February 13, 2019

8.1K
Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
07:27

Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase

Published on: April 29, 2010

13.6K

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • DNA replication necessitates the removal of numerous RNA primers from the lagging strand.
  • The coordination between DNA Polymerase III alpha (Pol IIIα), DNA Polymerase I (Pol I), and DNA ligase for primer removal is not fully understood.

Purpose of the Study:

  • To elucidate the coordination mechanism between enzymes involved in RNA primer removal during DNA replication.
  • To characterize the role of DNA Polymerase I in displacing and processing RNA primers.

Main Methods:

  • Cryogenic-electron microscopy (cryo-EM) was used to determine the structure of Pol I bound to a DNA substrate with RNA primers.
  • Biochemical assays were employed to analyze the enzymatic activities and substrate preparation for subsequent enzymes.

Main Results:

  • A four-point molecular handover mechanism was identified, where each enzyme's activity prepares the substrate for the next.
  • Cryo-EM revealed Pol I displaces RNA primers similarly to monomeric helicases.
  • The endonuclease domain of Pol I was shown to cut specifically at the RNA-DNA junction, facilitating ligation.

Conclusions:

  • The sequential action of Pol IIIα, Pol I polymerase, Pol I endonuclease, and DNA ligase ensures efficient RNA primer removal.
  • Pol I plays a crucial role in both primer displacement and preparing the DNA strand for ligation by LigA.