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Related Concept Videos

Phosphoinositides and PIPs01:42

Phosphoinositides and PIPs

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Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
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Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
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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,...
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PIWI-interacting RNAs, or piRNAs, are the most abundant short non-coding RNAs. More than 20,000 genes have been found in humans that code for piRNAs while only 2000 genes have been found for miRNAs. piRNAs can act at the transcriptional and post-transcriptional levels and have a vital role in silencing transposable elements present in germ cells. They are also involved in epigenetic silencing and activation. Previously, they were thought to function only in germ cells but new evidence suggests...
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Filopodia are thin, actin-rich cellular protrusions that play an important role in many fundamental cellular functions. They vary in their occurrence, length, and positioning in different cell types, suggesting their diverse roles.
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Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
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Related Experiment Video

Updated: Oct 9, 2025

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry
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Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

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Pif1 Activity is Modulated by DNA Sequence and Structure.

David G Nickens1, Matthew L Bochman1

  • 1Molecular & Cellular Biochemistry Department, Indiana University, Bloomington, Indiana 47405, United States.

Biochemistry
|December 21, 2021
PubMed
Summary

The Pif1 helicase binds structured DNA but unwinds unstructured DNA most efficiently. This suggests structured DNA inhibits Pif1

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Pif1 helicase is crucial for DNA metabolism, involved in processes like telomere length maintenance.
  • Its functions depend on DNA binding and ATP hydrolysis for unwinding DNA and secondary structures.
  • Previous studies explored Pif1's interaction with G-quadruplex DNA and ssDNA length requirements.

Purpose of the Study:

  • To investigate the effects of single-stranded DNA (ssDNA) length, sequence, and structure on Pif1's biochemical activities in vitro.
  • To characterize Pif1's ssDNA binding, ATPase, and helicase activities using various oligonucleotide substrates.

Main Methods:

  • Utilized recombinant, untagged Saccharomyces cerevisiae Pif1.
  • Performed in vitro biochemical assays including ssDNA binding, ATPase activity, and helicase activity assays.

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  • Employed a suite of oligonucleotide-based substrates to test Pif1 activity.

    • Main Results:

    • Pif1 preferentially binds to structured, G-rich ssDNA, but this binding did not maximally stimulate ATPase activity.
    • Significant DNA unwinding activity was observed at Pif1 concentrations as low as 250 pM.
    • Pif1 most efficiently unwinds DNA fork substrates with unstructured ssDNA tails.

    Conclusions:

    • Structured DNA may inhibit Pif1's conformational changes required for ATP hydrolysis-coupled DNA translocation and unwinding.
    • Pif1's efficiency in unwinding DNA is dependent on the structure of the ssDNA substrate.
    • These findings provide insights into the regulation of Pif1 helicase activity.