Ribosome movement on its template transcript plays an important role in mRNA quality control and translational regulation. Surprisingly, some regulatory nascent polypeptides stall their own translation when they are still inside the ribosome tunnel, a path that every newly synthesized protein traverses to leave the large ribosomal subunit. Despite the universality and structural conservation of the tunnel, its function is still rather mysterious. It is unclear how the tunnel deciphers the arrest signals and discriminates non-regulatory peptides from regulatory peptides. During the K99 phase I used E. coli SecM as a model to investigate how the arrest sequence motif is recognized inside the tunnel and how the timing of the release of arrest is achieved. In the ROO phase, we aim to advance our understanding of translatlonal attenuation by investigating the biological activities of the ribosome tunnel by characterizing the regulatory role of small peptides and by identifying peptide sequences within larger proteins that promote ribosome stalling/pausing. Specifically we will use an Integrative approach that combines comparative proteomics, bacterial genetics, biochemical methods and genome-wide ribosome profiling to elucidate the fundamental principles of these processes. This work will not only provide new information on the molecular responses of the tunnel to disparate nascent chains, but will also offer a broader view of the diversity of translational control systems and the translation machinery that is conserved in all living systems.

Public Health Relevance

The ribosome plays a universally conserved role in catalyzing protein synthesis in all living organisms. Misregulation of translation appears to be associated with a variety of human diseases. Results from this work will provide detailed molecular insight into the mechanisms of translational regulation and may thereby lead to potential therapies to control the misregulated processes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Transition Award (R00)
Project #
5R00GM094212-03
Application #
8393462
Study Section
Special Emphasis Panel (NSS)
Program Officer
Bender, Michael T
Project Start
2011-12-05
Project End
2014-11-30
Budget Start
2012-12-01
Budget End
2013-11-30
Support Year
3
Fiscal Year
2013
Total Cost
$234,064
Indirect Cost
$69,980
Name
Saint Louis University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
050220722
City
Saint Louis
State
MO
Country
United States
Zip Code
63103
Davis, Amber R; Gohara, David W; Yap, Mee-Ngan F (2014) Sequence selectivity of macrolide-induced translational attenuation. Proc Natl Acad Sci U S A 111:15379-84
Yap, Mee-Ngan F; Bernstein, Harris D (2013) Mutations in the Escherichia coli ribosomal protein L22 selectively suppress the expression of a secreted bacterial virulence factor. J Bacteriol 195:2991-9
Yap, Mee-Ngan F (2013) The double life of antibiotics. Mo Med 110:320-4