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Determination

Determination

During embryogenesis, cells become progressively committed to different fates through a two-step process: specification followed by determination. Specification is demonstrated by removing a segment of an early embryo, “neutrally” culturing the tissue in vitro—for example, in a petri dish with simple medium—and then observing the derivatives. If the cultured region gives rise to cell types that it would normally generate in the embryo, this means that it is specified. In contrast, determination...

Nucleosome Remodeling

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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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Eukaryotic cells have specialized enzymes called ATP-dependent nucleosome remodeling enzymes. These enzymes...

DNA Damage can Stall the Cell Cycle

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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...

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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...

Nucleotide Excision Repair

Nucleotide Excision Repair

DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...

DNA Damage Can Stall the Cell Cycle

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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...

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Related Experiment Video

Updated: Jul 15, 2026

Double Whole Mount in situ Hybridization of Early Chick Embryos

Double Whole Mount in situ Hybridization of Early Chick Embryos

Published on: October 27, 2008

rasiRNAs, DNA damage, and embryonic axis specification.

W E Theurkauf¹, C Klattenhoff, D P Bratu

¹Program in Molecular Medicine and Program in Cell Dynamics, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.

Cold Spring Harbor Symposia on Quantitative Biology

|March 27, 2007

Summary

Drosophila repeat-associated small interfering RNAs (rasiRNAs) normally silence transposons. Mutations disrupt axis specification, but ATR/Chk2 kinases suppress these defects by preventing DNA damage responses in the germ line.

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Double Whole Mount in situ Hybridization of Early Chick Embryos

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Published on: October 27, 2008

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Detection of DNA Double-Stranded Breaks in Mouse Oocytes

Detection of DNA Double-Stranded Breaks in Mouse Oocytes

Published on: June 23, 2023

Area of Science:

Developmental Biology
RNA Biology
Genetics

Background:

Drosophila repeat-associated small interfering RNAs (rasiRNAs) are crucial for silencing retrotransposons and specific genetic loci.
Mutations in key rasiRNA pathway genes (armitage, spindle-E, aubergine) cause severe defects in embryonic axis specification during oogenesis, including disrupted microtubule organization and mRNA localization.

Purpose of the Study:

To investigate the role of DNA damage response kinases (ATR and Chk2) in suppressing rasiRNA pathway-mediated developmental defects.
To elucidate the mechanism by which rasiRNA pathway mutations lead to axis specification failure.

Main Methods:

Genetic analysis of rasiRNA pathway mutants and DNA damage response mutants (mei-41, mnk) in Drosophila.
Assessment of cytoskeletal organization, mRNA localization (osk, grk), and DNA double-strand break markers (gamma-H2Av foci).
Analysis of protein phosphorylation (Vasa) in response to rasiRNA pathway mutations.

Main Results:

Mutations in mei-41 (ATR) and mnk (Chk2) significantly suppressed cytoskeletal and RNA localization defects caused by rasiRNA pathway mutations.
Stellate and retrotransposon silencing were not restored in mei-41 and mnk double mutants.
rasiRNA pathway mutations induced germ-line-specific gamma-H2Av foci, indicating DNA double-strand breaks.
armitage mutations resulted in Chk2-dependent Vasa phosphorylation.

Conclusions:

The Drosophila rasiRNA pathway plays a role in suppressing DNA damage within the germ line.
Disruptions in the rasiRNA pathway activate an ATR/Chk2-dependent DNA damage response, which impairs microtubule polarization and mRNA localization, leading to failed axis specification.