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Related Concept Videos

Protein Complex Assembly02:41

Protein Complex Assembly

Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
Protein Complex Assembly02:41

Protein Complex Assembly

Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
Assembly of Signaling Complexes01:30

Assembly of Signaling Complexes

Multiprotein signaling complexes are formed in a dynamic process involving protein-protein interactions at the cytoplasmic domain of transmembrane receptors or enzymatic and non-enzymatic proteins associated with the receptor. These complexes ensure the activation and propagation of intracellular signals that regulate cell functions.
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Coat Assembly and GTPases01:33

Coat Assembly and GTPases

Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
Microtubule Formation01:23

Microtubule Formation

Microtubules are dynamic structures that undergo continuous assembly and disassembly. They originate from specialized multi-protein complexes known as microtubule organizing centers or MTOCs. Within the MTOC, the point of origin of the microtubule is known as the minus end, while the end radiating outward is the plus end. Microtubules serve two primary functions — the organization of spindle complexes to separate sister chromatids during mitotic or meiotic cell division and the formation of...
Replication in Eukaryotes01:29

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...

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Related Experiment Video

Updated: Jul 12, 2026

Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes
08:44

Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes

Published on: October 23, 2014

ATP-dependent assembly of the human origin recognition complex.

Khalid Siddiqui1, Bruce Stillman

  • 1Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.

The Journal of Biological Chemistry
|August 25, 2007
PubMed
Summary

Human origin recognition complex (ORC) assembly requires ATP binding and is cell cycle-regulated. ORC disassembles during S phase, suggesting a unique mechanism for restricting DNA replication.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • The origin recognition complex (ORC) is crucial for initiating DNA replication in eukaryotes.
  • ORC binds autonomously replicating sequences in an ATP-dependent manner.
  • Understanding human ORC assembly and regulation is key to cell cycle control.

Purpose of the Study:

  • To clone and express human ORC subunits as recombinant proteins.
  • To investigate the role of ATP binding in human ORC assembly.
  • To determine the cell cycle regulation of human ORC.

Main Methods:

  • Recombinant protein expression and purification of human ORC subunits.
  • In vitro assembly assays assessing ATP-binding dependence.
  • Immunofluorescence and immunoprecipitation for cell cycle analysis.

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Examining Proteasome Assembly with Recombinant Archaeal Proteasomes and Nondenaturing PAGE: The Case for a Combined Approach

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Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique
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Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

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Related Experiment Videos

Last Updated: Jul 12, 2026

Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes
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Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes

Published on: October 23, 2014

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Examining Proteasome Assembly with Recombinant Archaeal Proteasomes and Nondenaturing PAGE: The Case for a Combined Approach

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Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique
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Main Results:

  • Human ORC assembly in vitro is ATP-binding dependent for Orc4 and Orc5, but not Orc1.
  • ORC associates with a nuclear structure in a cell cycle-dependent manner.
  • ORC disassembles during S phase, and Orc6 interacts with Orc3 within the complex.

Conclusions:

  • Human ORC assembly is uniquely regulated by ATP binding.
  • Cell cycle-dependent disassembly of ORC contributes to the once-per-cell-cycle DNA replication.
  • These findings offer insights into the precise control of DNA replication initiation.