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molecular biology, cellular biology, biochemistry, microscopy

WHAT WE DO

Key Points:

  • Elucidate the mechanism of ribosomal protein transport and assembly.

  • Investigate the role of sugar metabolism on protein synthesis.

Detailed Description:

Proteins are biological molecules that provide structural support to the cell and enable its functions. They can work individually or bundle together with other molecules to form complexes with specific functions. Proteins and other materials move to their sites of action via cellular transport with the help of transporters, carriers, motor proteins, or vesicles. Intracellular transport ensures that constituents reach their final intended destination, which is critical to the growth and life of the cell.

 

Ribosomes are the cellular machines responsible for making proteins in all organisms. They consist of two subunits made up of RNA molecules and multiple ribosomal proteins, which are translocated across three different compartments: the cytoplasm, the nucleus, and the nucleolus.

 

The FIRE CellEx stream investigates how the modification of ribosomes controls the transport of these machines between different compartments of cells in a manner that will expand fundamental knowledge of cellular biology and impact a number of human diseases.

WHY IT MATTERS

Key Points:

  • Defects in ribosome biogenesis underlie genetic diseases and cancer development.

Detailed Description:

Mutations in ribosomal protein genes or disruption of ribosome production cause Diamond Blackfan anemia (DBA), Shwachman Diamond syndrome (SDS), X-linked dyskeratosis congenita (DC), and other genetic disorders, characterized by bone marrow failure, developmental anomalies, defective immunity, and skeletal malformations, among others.

 

These tissue-specific pathologies, compiled under the term ribosomopathies, have no known cure and are often fatal. Changes in ribosome function and protein synthesis are associated with the progression of solid tumors and hematopoietic oncogenic transformation. Therapeutic approaches that target ribosome biogenesis and function can benefit patients with congenital ribosomal disorders and related malignancies.

 

The FIRE CellEx research comprehensively analyses the ribosome assembly landscape, identifying steps susceptible to interventions and providing mechanistic insights into potential treatments.  

WHAT YOU LEARN

Key Points:

  • To dissect the roadmap for ribosome subunit building using cell biology, biochemistry, and molecular biology techniques.

Detailed Description:

The experimental strategy for the FIRE CellEx stream includes in vitro, ex vivo, and in vivo approaches to track the glycosylation dynamics of individual ribosomal proteins as they integrate into molecular complexes.

 

Students learn various methods, including cell culture, molecular cloning, DNA recombinant technology, protein expression and affinity purification, site-directed mutagenesis, and glycosyltransferase reactions.

 

Protein-sugar modifications are detected by click chemistry, protein separation, and immunodetection and confirmed with mass spectrometry.

 

Ribosome assembly intermediates are extracted using subcellular fractionation and density-gradient centrifugation and visualized within cells by live fluorescence microscopy.

 

Genome-wide effects of aberrant sugar-ribosome metabolism on translation are monitored by ribosome profiling with implications for human disease.  

Related Resources

More information soon.