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Related Concept Videos

Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to form...
Lampbrush Chromosomes01:51

Lampbrush Chromosomes

In 1882, Flemming observed lampbrush chromosomes (LBC) in salamander eggs. Later in 1892, Rückert observed LBCs in shark egg cells and coined the term "lampbrush chromosomes" because they looked like brushes used to clean kerosene lamps.
LBCs are made up of two pairs of conjugating homologous chromatids. Each chromatid consists of alternatively positioned regions of condensed-inactive chromatin and loosely placed-active side loops, which can be contracted and extended. The loops resemble the...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
Genomic DNA in Eukaryotes00:58

Genomic DNA in Eukaryotes

Eukaryotes have large genomes compared to prokaryotes. To fit their genomes into a cell, eukaryotic DNA is packaged extraordinarily tightly inside the nucleus. To achieve this, DNA is tightly wound around proteins called histones, which are packaged into nucleosomes that are joined by linker DNA and coil into chromatin fibers. Additional fibrous proteins further compact the chromatin, which is recognizable as chromosomes during certain phases of cell division.
Euchromatin01:01

Euchromatin

The extent of chromatin compaction can be studied by staining chromatin using specific DNA binding dyes. Under the microscope, the dense-compacted regions take up more dye, appearing darker, while the less-compact areas take up less dye and appear lighter. Based on the compaction level, chromatins are classified into two primary forms – euchromatin and heterochromatin.
Euchromatin is the less dense region of the chromatin and stains lighter. Euchromatin contains histone H3 extensively...

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Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans
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Eukaryotic Argonautes come into focus.

Claus-D Kuhn1, Leemor Joshua-Tor

  • 1W. M. Keck Structural Biology Laboratory, Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.

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Structural insights into human Argonaute-2 (hAgo2) and yeast Argonaute provide a molecular basis for understanding RNA-induced silencing complex (RISC) function. These advances in RNA interference (RNAi) pave the way for detailed mechanistic studies of eukaryotic gene silencing.

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Genetics

Background:

  • Small RNAs regulate gene expression through RNA-induced silencing complex (RISC).
  • Eukaryotic Argonaute structures were previously unavailable, limiting detailed mechanistic studies of RISC.
  • Different small RNA classes utilize distinct biogenesis pathways but converge on Argonaute association.

Purpose of the Study:

  • To review recent structural advancements in eukaryotic Argonautes.
  • To highlight how these structures elucidate RNA-induced silencing complex (RISC) mechanisms.
  • To provide a foundation for future molecular studies of RNA interference (RNAi).

Main Methods:

  • X-ray crystallography of human Argonaute-2 (hAgo2) in complex with guide RNA.
  • Determination of hAgo2 structure in complex with a specific microRNA (miRNA).
  • Structural analysis of a budding yeast Argonaute.

Main Results:

  • The structure of hAgo2 was determined in complex with heterogeneous guide RNA.
  • The structure of hAgo2 was solved in complex with a specific miRNA.
  • The structure of a budding yeast Argonaute was elucidated.

Conclusions:

  • Recent Argonaute structures provide unprecedented molecular detail on RISC.
  • These structural insights enable a deeper mechanistic understanding of eukaryotic RNA interference (RNAi).
  • Future research can now explore RISC function at a molecular level.