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Related Concept Videos

Telomeres and Telomerase02:41

Telomeres and Telomerase

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In eukaryotic DNA replication, a single-stranded DNA fragment remains at the end of a chromosome after the removal of the final primer. This section of DNA cannot be replicated in the same manner as the rest of the strand because there is no 3’ end to which the newly synthesized DNA can attach. This non-replicated fragment results in gradual loss of the chromosomal DNA during each cell duplication. Additionally, it can induce a DNA damage response by enzymes that recognize single-stranded...
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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Restarting Stalled Replication Forks02:37

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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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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
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During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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CST-polymerase α-primase solves a second telomere end-replication problem.

Hiroyuki Takai1, Valentina Aria2, Pamela Borges1

  • 1Laboratory for Cell Biology and Genetics, Rockefeller University, New York, NY, USA.

Nature
|February 28, 2024
PubMed
Summary
This summary is machine-generated.

Telomere maintenance involves two replication problems. Telomerase handles G-rich strand shortening, while the Ctc1-Stn1-Ten1 polymerase-primase complex resolves C-rich strand issues during DNA replication.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Telomeres protect chromosome ends from degradation and fusion.
  • Telomere shortening occurs due to the end-replication problem during DNA synthesis.
  • Telomerase counteracts shortening of the G-rich strand at telomeres.

Purpose of the Study:

  • To identify and characterize a second end-replication problem affecting the C-rich telomeric strand.
  • To elucidate the mechanism and factors involved in resolving C-strand replication issues.
  • To understand the combined roles of telomerase and CST-Polα-primase in telomere maintenance.

Main Methods:

  • In vitro DNA replication assays using telomeric DNA substrates.
  • Analysis of telomere length and C-strand integrity in cells lacking CST-Polα-primase.
  • Quantitative measurement of telomeric repeat loss per cell division.

Main Results:

  • Lagging-strand DNA synthesis stalls approximately 26 nt from the telomere end, leaving a C-strand gap.
  • CST-Polα-primase mediates fill-in synthesis to resolve this C-strand replication defect.
  • Cells lacking CST-Polα-primase exhibit significant C-strand shortening at both leading and lagging telomere ends.

Conclusions:

  • Canonical DNA replication presents two end-replication challenges: G-strand loss and C-strand incomplete synthesis.
  • Telomerase maintains the G-rich strand, while CST-Polα-primase is essential for C-strand maintenance.
  • Proper telomere length requires coordinated action of telomerase and CST-Polα-primase to address both replication problems.