molecular biology, cellular biology, biochemistry, microscopy

WHAT WE DO

Key Points:

Explain how ribosomal proteins are transported and assembled inside the cell.
Investigate the role of various chemical modifications on proteins that participate in translation.

Detailed Description:

Human cells contain internal compartments called organelles that fulfill specific activities, similar to the body's organs. Proteins and other molecules inside the cell constantly move across these different compartments to carry out their functions. While traveling, proteins often change their chemical composition and connect with other molecules to help them move through the cell and reach their destination.

Among the molecules in constant motion are the ribosomes, essential to decoding the information in genes and translating it into proteins. In human cells, eighty unique ribosomal proteins move across different cell compartments and assemble with RNA molecules to make a functional ribosome. The FIRE CellEx stream investigates the chemical modifications and molecular interactions each ribosomal protein encounters during its transport inside the cell before being incorporated into a ribosome.

WHY IT MATTERS

Key Points:

Defects in ribosomal protein function cause genetic diseases and cancer progression.

Detailed Description:

Mutations in ribosomal proteins cause rare genetic diseases and promote cancer development, especially leukemia. Rare ribosomal disorders, such as Diamond Blackfan Anemia (DBA), Shwachman Diamond syndrome (SDS), and X-linked dyskeratosis congenita (DC), are characterized by bone marrow failure, congenital disabilities, and severe immune alterations. Most ribosomal diseases have no known cure and are often fatal.

Developing biomedical approaches targeting ribosomal protein activity could benefit patients with rare congenital diseases and cancer. The FIRE CellEx research analyses the ribosome assembly landscape, identifying steps susceptible to therapeutic interventions and providing insights into potential disease treatments.

WHAT YOU LEARN

Key Points:

To dissect the ribosomal protein transport roadmap using cell biology, biochemistry, and molecular biology techniques.

Detailed Description:

The experimental strategy for FIRE CellEx includes in vitro and in vivo approaches to follow the dynamics of individual ribosomal proteins as they integrate into molecular assemblies.

In FIRE Semester 2, students focus on molecular biology techniques, including primer design, polymerase chain reaction (PCR), DNA agarose gel electrophoresis, molecular cloning, bacterial transformation, plasmid DNA purification, and sequencing alignment.

In FIRE Semester 3, we focus on protein biochemistry, using methods for protein expression (in bacteria, cell-free systems, and cells in culture), protein separation by acrylamide gel electrophoresis, affinity purification by chromatography and immunoprecipitation, and Western blots. We are working on optimizing other experimental strategies, such as subcellular fractionation, centrifugation, and fluorescence microscopy.

In addition to wet lab techniques, students learn to apply the scientific method, collaborate in teams, and communicate scientific findings through poster and oral presentations at scientific meetings.