Rapidly dividing cells must produce 2,000 new ribosomes every minute and ensure that they are fully functional. The mRNA entry channel is formed on the beak structure of the small ribosomal subunit, and two out of the three proteins at the beak are associated with DBA, an inherited human disease, caused by haploinsufficiency of ribosomal proteins (r-proteins). As expected from loss of r-proteins, patients have low cell numbers in rapidly dividing tissues, but are also predisposed to cancer, suggesting that these patients fail in producing both the numbers and quality of ribosomes needed. Our recent data indicates that r- proteins at the mRNA entry channel are assembled late during maturation, and that assembly is regulated by release of the assembly factor Ltv1, which requires phosphorylation by the Hrr25 kinase. The overall goal of this project is to dissect the assembly of the mRNA entry channel using structural and biochemical methods.
In Aim 1 we will dissect how Ltv1 dissociation affects the structure of pre-40S subunits, and the mRNA binding channel. We will also test how release of Ltv1 is linked to recruitment of the ribosomal protein Asc1, which is required for translation of IRES-containing viral messages.
In Aim 2 we will use structural and yeast biochemical methods to define how incorporation of Rps10 at the beak is linked to 18S rRNA processing at the platform, via a network of proteins involved in DBA. These studies will provide insight into assembly events that are very rapid within cells and increase our understanding of the mechanisms by which defects in ribosome maturation can lead to tumorigenesis.

Public Health Relevance

Rapidly dividing cells must produce 2,000 new ribosomes every minute and ensure that they are fully functional and not prone to mistranslation. Diamond Blackfan Anemia (DBA) and 5q- syndrome are human diseases caused by haploinsufficiency of ribosomal proteins (r-proteins). As expected from loss of r- proteins, patients have low cell numbers in rapidly dividing tissues but are also predisposed to cancer, suggesting that these patients fail in producing both the numbers and quality of ribosomes needed. The mRNA entrance channel is formed by the beak structure of the small subunit, and two out of the three proteins at the beak are associated with DBA. In this work we will build on our previously developed tools and combine biochemical and structural methods to dissect the mechanisms by which beak assembly is facilitated and regulated. The insights from this work will also shed light into the ways by which healthy cells guard against the release of ribosomes with incompletely assembled beak structures into the translating pool, thereby protecting against tumorigenesis.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01GM117093-02
Application #
9197652
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Flicker, Paula F
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Scripps Florida
Department
Type
DUNS #
148230662
City
Jupiter
State
FL
Country
United States
Zip Code
33458