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Related Concept Videos

Animal Mitochondrial Genetics02:59

Animal Mitochondrial Genetics

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Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
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Mitochondrial Protein Sorting01:39

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Mitochondria are double-membrane organelles of the eukaryotes involved in cellular metabolism, signaling, ATP synthesis, and programmed cell death.  Each of these processes requires specific proteins and enzymes that must be correctly sorted to the right mitochondrial subcompartment for the proper functioning of the organelle.
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Mitochondrial Precursor Proteins01:39

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Mitochondrial precursors are partially unfolded or loosely folded polypeptide chains. Newly synthesized precursors are inhibited from spontaneously folding into their native conformation by the cytosolic chaperones, heat shock proteins 70 (Hsp70), and mitochondrial import stimulation factors (MSFs). Precursors bound to MSFs are guided to the TOM70-TOM37 receptors, while precursors bound to Hsp70  chaperones are targetted to TOM20-TOM22 receptor complexes.
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Translocation of Proteins into the Mitochondria01:19

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Mitochondrial precursors are translocated to the internal subcompartments via independent mechanisms involving distinct protein machineries called translocases.
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Replication in Eukaryotes01:29

Replication in Eukaryotes

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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.
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Protein Transport into the Inner Mitochondrial Membrane01:34

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Nuclear encoded mitochondrial precursors are imported to the inner membrane in a multistep process involving two separate translocons, TIM22 and TIM23. TIM23 is a cation-selective pore that remains closed by the N terminal segment of the protein. Negative charges on the TIM23 act as a receptor for the incoming precursor, pulling the positively charged matrix-targeting sequence for peptide insertion and translocation.
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Related Experiment Video

Updated: Aug 9, 2025

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA
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5'-End Mapping in Human Mitochondrial DNA.

Andranik Durgaryan1, Anders R Clausen2

  • 1Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.

Methods in Molecular Biology (Clifton, N.J.)
|February 22, 2023
PubMed
Summary

We developed 5'-End-sequencing (5'-End-seq) to map DNA 5'-ends genome-wide using next-generation sequencing. This assay reveals insights into DNA integrity, replication, and processing events across the entire genome.

Keywords:
5′-End-seqBioinformaticsDNA polymeraseDNA replicationGenomicsNext-generation sequencingRNase H1mtDNA

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biochemistry

Background:

  • Understanding DNA dynamics is crucial for cellular health.
  • Existing methods may have limitations in comprehensively mapping DNA 5 eal;-ends genome-wide.
  • Mitochondrial DNA (mtDNA) integrity and replication are key areas of research.

Purpose of the Study:

  • To introduce a novel assay, 5 eal;-End-sequencing (5 eal;-End-seq), for high-resolution mapping of DNA 5 eal;-ends.
  • To apply this method to study free 5 eal;-ends in mitochondrial DNA (mtDNA).
  • To demonstrate the utility of 5 eal;-End-seq for investigating genome-wide DNA processing events.

Main Methods:

  • Development of a next-generation sequencing assay named 5 eal;-End-sequencing (5 eal;-End-seq).
  • Application of 5 eal;-End-seq on Illumina sequencing platforms.
  • Isolation of mitochondrial DNA (mtDNA) from fibroblast cells for analysis.

Main Results:

  • Successful mapping of free 5 eal;-ends across the genome using 5 eal;-End-seq.
  • Demonstration of the assay's capability to analyze mtDNA.
  • Identification of potential applications in studying DNA integrity and processing.

Conclusions:

  • 5 eal;-End-sequencing is a powerful new tool for genome-wide 5 eal;-end mapping.
  • The method provides insights into DNA replication, integrity, and processing mechanisms.
  • This assay has broad applications in various fields of genomic research.