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Related Concept Videos

DNA Helicases00:55

DNA Helicases

DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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Nucleosomes are the basic units of chromatin compaction. Each nucleosome consists of the DNA bound tightly around a histone core, which makes the DNA inaccessible to DNA binding proteins such as DNA polymerase and RNA polymerase. Hence, the fundamental problem is to ensure access to DNA when appropriate, despite the compact and protective chromatin structure.
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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...
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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
DNA Damage Can Stall the Cell Cycle02:36

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Allosteric Proteins-ATCase

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CD Spectroscopy to Study DNA-Protein Interactions
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ATP binding and hydrolysis by Mcm2 regulate DNA binding by Mcm complexes.

Brent E Stead1, Catherine D Sorbara, Christopher J Brandl

  • 1Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, Ontario, Canada.

Journal of Molecular Biology
|June 23, 2009
PubMed
Summary

Minichromosome maintenance (Mcm) proteins regulate DNA replication. Nucleotide binding by Mcm2 is critical for controlling the DNA binding and unwinding activities of Mcm complexes, impacting yeast viability.

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Published on: March 31, 2010

Area of Science:

  • Molecular Biology
  • Eukaryotic DNA Replication
  • Protein Biochemistry

Background:

  • The minichromosome maintenance (Mcm) complex (Mcm2-7) is the core eukaryotic replicative helicase.
  • Subcomplexes, such as Mcm4/6/7, are capable of DNA unwinding.
  • Previous research indicated Mcm2 inhibits Mcm4/6/7 DNA unwinding activity.

Purpose of the Study:

  • To investigate the role of nucleotide binding by Saccharomyces cerevisiae Mcm2 in regulating Mcm complex DNA binding and unwinding.
  • To determine if Mcm2 requires nucleotide binding for its inhibitory function on Mcm4/6/7.

Main Methods:

  • Utilized an Mcm4/6/7 subcomplex as a tool to study Mcm2 regulation.
  • Employed an Mcm2 mutant defective for ATP hydrolysis (K549A) and ATP analogues.
  • Assessed DNA binding and unwinding activities of Mcm complexes under various nucleotide conditions.

Main Results:

  • Mcm2 interaction with Mcm4/6/7 is insufficient for inhibition; nucleotide binding by Mcm2 is required.
  • ADP binding to Mcm2 is necessary to inhibit DNA binding and unwinding by Mcm4/6/7, independent of Mcm3/5.
  • The Mcm2(K549A) mutation significantly altered DNA binding properties of the Mcm2-7 complex and impaired yeast viability.

Conclusions:

  • The nucleotide-bound state of Mcm2 is critical for regulating the DNA binding and unwinding activities of Mcm4/6/7 and Mcm2-7 complexes.
  • ATP hydrolysis by Mcm2 plays a vital role in the regulation of native Mcm complexes.
  • Mcm2's nucleotide-dependent regulation is essential for yeast viability.