Related Experiment Video

Updated: May 13, 2026

Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates
14:32

Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates

Published on: February 27, 2016

Human SIRT1 regulates DNA binding and stability of the Mcm10 DNA replication factor via deacetylation

Samuel T Fatoba1, Silvia Tognetti, Melissa Berto

  • 1Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK.

Nucleic Acids Research
|March 2, 2013
PubMed

Insights

SIRT1 deacetylates human Mcm10, a key DNA replication factor, enhancing its stability and DNA binding. This interaction is crucial for DNA replication initiation and progression, linking metabolism and DNA synthesis.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Mcm10 is a eukaryotic DNA replication initiation factor essential for replisome assembly and function.
  • Human Mcm10 possesses two DNA-binding domains: the conserved internal domain (ID) and the metazoan-specific C-terminal domain (CTD).
  • SIRT1 is an NAD-dependent deacetylase involved in metabolism, longevity, gene expression, and genomic stability.

Purpose of the Study:

  • To investigate the regulatory relationship between human Mcm10 and SIRT1.
  • To elucidate the role of SIRT1-mediated deacetylation in Mcm10 function and stability.
  • To explore the impact of Mcm10 acetylation on its DNA-binding domains and replication activity.

Main Methods:

  • In vivo and in vitro binding assays to confirm Mcm10-SIRT1 interaction.
  • Deacetylation assays to assess SIRT1's enzymatic activity on Mcm10.
  • Analysis of Mcm10 stability and DNA-binding affinity upon SIRT1 modulation.
  • Investigation of the distinct effects of acetylation/deacetylation on Mcm10's ID and CTD.

Main Results:

  • Human Mcm10 is acetylated and its acetylation is regulated by SIRT1.
  • SIRT1 physically binds and deacetylates Mcm10 both in vivo and in vitro.
  • SIRT1 deacetylation modulates Mcm10 stability and its ability to bind DNA.
  • Mcm10 and SIRT1 act synergistically in DNA replication fork initiation.
  • Acetylation/deacetylation distinctly affects Mcm10's internal and C-terminal DNA-binding domains.

Conclusions:

  • Protein acetylation plays a significant role in DNA replication initiation and progression.
  • SIRT1 regulates Mcm10 activity through deacetylation, impacting DNA replication.
  • This study suggests a crosstalk between metabolic regulation (via SIRT1) and DNA synthesis (via Mcm10).

More Related Videos

Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique09:14

Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique

Published on: January 14, 2016

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images09:42

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images

Published on: September 7, 2017

Related Concept Videos

Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.
Spreading of Chromatin Modifications02:25

Spreading of Chromatin Modifications

The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer is an enzyme that can...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
Histone Modification02:32

Histone Modification

The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression.
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...