Ribosome assembly is an evolutionarily conserved and energy intensive process required for cellular growth, proliferation, and maintenance. In yeast, assembly of the small ribosomal subunit (SSU) requires approximately 75 assembly factors that act in coordination to form the SSU processome, a 6 MIDa ribonucleoprotein complex. These factors are responsible for the formation of the SSU processome, but are not a part of the mature ribosome. The SSU processome is required for processing, modifying, and folding the pre-ribosomal RNA (rRNA) to prepare it for incorporation into the mature SSU. Although the protein composition of the SSU processome has been known for some time, the interaction network of the proteins required for its assembly and function has remained poorly defined. Here, we have used a semi-high-throughput yeast two-hybrid (Y2H) assay and co-immunoprecipitation validation method to produce a high-confidence interactome of SSU processome assembly factors (SPAFs), providing essential insight into SSU assembly and ribosome biogenesis. Further, we used glycerol density gradient sedimentation to reveal the presence of putative protein subcomplexes that have not previously been observed. Our work not only provides essential insight into SSU assembly and ribosome biogenesis, but also serves as an important resource for future investigations into how defects in SSU biogenesis and assembly cause congenital disorders of ribosomes known as ribosomopathies.
|Advisor:||Baserga, Susan J.|
|School Location:||United States -- Connecticut|
|Source:||DAI-B 79/05(E), Dissertation Abstracts International|
|Keywords:||Interactome, Ribosome Biogenesis, SSU Processome, Saccharomyces Cerevisiae, Small Subunit (SSU), Yeast Two-Hybrid (Y2H)|
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