Comprehensive Analysis Of Post-Translational Modifications (PTMs) In Proteins Involved In Histone Pre-mRNA Processing By High Performance nanoLC-MS
EMSL Project ID
30458
Abstract
All eukaryotic mRNAs undergo a series of posttranscriptional processing events that lead to the generation of a mature mRNA transcript which is then translated. These pre-mRNA processing steps include 3 end formation, 5 capping, splicing, and editing. Correct processing requires the specific assembly of protein and RNA components. The role of posttranslational modifications in RNA assembly and in modulating protein function is not understood. RNA binding proteins are known to be posttranslationally modified by phosphorylation, lysine acetylation, arginine methylation, and sumoylation.The focus of my research is to understand how the histone pre-mRNA processing complex assembles and to gain insight into the role of posttranslational modifications in its assembly. In particular, we have been studying the histone mRNA specific factor Stem-Loop Binding Protein (SLBP) and its interaction with histone mRNA. SLBP is a phosphoprotein that is phosphorylated at a number of Ser, Thr residues only two of which have been characterized. It is likely that it is also modified at Arg and Lys residues and could be sumoylated. This proposal is focused on gaining insight into the sites of modification that occur in human and Drosophila SLBP in baculovirus-expressed SLBP as well as SLBP isolated from intact cells. Our long-term goal is to characterize the modification state of the protein as a function of the cell cycle, thereby gaining molecular insight into how the protein functions in vivo. NanoLC-MS would be particularly suitable in these efforts. The information obtained from these studies would provide new insight into how genes are controlled and regulated and also could aid in the design of novel inhibitors in cancer therapy that target the SLBP-RNA complex.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2008-09-30
End Date
2009-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members