Posttranslational modification of proteins in signal pathways is one of the most pivotal control points in vivo for the activation, inactivation, and degradation of factors governing nearly all physiological processes. The goal of this proposal is to define the molecular mechanism and in vivo functions of protein arginylation, a very poorly understood post-translational modification that affects an estimated 25% of the cellular proteome. Arginylation plays a key role in cell migration and developmental morphogenesis, and is implicated in major events of cell metabolism, physiology, and human disease. Our preliminary data strongly suggest that arginylation regulates its protein targets through rapid and reversible modulation of their activity and protein interactions and constitutes a truly essential and dynamic biological regulator, however virtually nothing is known about the substrate site specificity and the molecular mechanisms of this reaction. This proposal will address the mechanisms of the arginyl transfer enzyme (ATE1) and the regulation of its substrate specificity through an integrated approach that will provide the first mechanistic insights into this enigmatic posttranslational modification. We propose the following specific aims: (1) To determine site specificity and properties of N- terminal arginylation; (2) To test the mechanisms of arginylation at internal protein sites in intact proteins; and (3) To test the effect of cellular components on he rate and site specificity of arginylation by different ATE1 isoforms and modulate its activity in vivo. Together, the proposed studies will elucidate the molecular mechanism of a novel posttranslational modification with major biological significance. These studies will ultimately enable differential modulation of ATE1 activity and biological targeting, essential for exploring is therapeutic potential in critical arginylation-dependent human conditions, including heart disease, cancer, neurodegeneration, and aging. The results of our studies will open new possibilities of functional arginylation analysis and targeted manipulation of arginylation of key proteins during essential processes in normal physiology and disease.

Public Health Relevance

Posttranslational modifications play a key role in protein functions, and their abnormalities lead to severe disease states, including heart disease and cancer - the two leading causes of death in the United States. The current proposal will delineate the mechanistic basis for arginylation, a novel posttranslational modification implicated in heart disease and cancer, with the goal of enabling the development of new therapeutics that modulate different arginylation-dependent mechanisms in disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
4R01GM104003-04
Application #
9068168
Study Section
Macromolecular Structure and Function E Study Section (MSFE)
Program Officer
Gerratana, Barbara
Project Start
2013-09-01
Project End
2017-05-31
Budget Start
2016-06-01
Budget End
2017-05-31
Support Year
4
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Pavlyk, Iuliia; Leu, Nicolae A; Vedula, Pavan et al. (2018) Rapid and dynamic arginylation of the leading edge ?-actin is required for cell migration. Traffic 19:263-272
Wang, Junling; Pejaver, Vikas Rao; Dann, Geoffrey P et al. (2018) Target site specificity and in vivo complexity of the mammalian arginylome. Sci Rep 8:16177
Wang, Junling; Pavlyk, Iuliia; Vedula, Pavan et al. (2017) Arginyltransferase ATE1 is targeted to the neuronal growth cones and regulates neurite outgrowth during brain development. Dev Biol 430:41-51
Efimova, Nadia; Korobova, Farida; Stankewich, Michael C et al. (2017) ?III Spectrin Is Necessary for Formation of the Constricted Neck of Dendritic Spines and Regulation of Synaptic Activity in Neurons. J Neurosci 37:6442-6459
Wang, Junling; Han, Xuemei; Leu, Nicolae Adrian et al. (2017) Protein arginylation targets alpha synuclein, facilitates normal brain health, and prevents neurodegeneration. Sci Rep 7:11323
Leite, Felipe S; Minozzo, Fábio C; Kalganov, Albert et al. (2016) Reduced passive force in skeletal muscles lacking protein arginylation. Am J Physiol Cell Physiol 310:C127-35
Rai, R; Zhang, F; Colavita, K et al. (2016) Arginyltransferase suppresses cell tumorigenic potential and inversely correlates with metastases in human cancers. Oncogene 35:4058-68
Zhang, Fangliang; Patel, Devang M; Colavita, Kristen et al. (2015) Arginylation regulates purine nucleotide biosynthesis by enhancing the activity of phosphoribosyl pyrophosphate synthase. Nat Commun 6:7517
Kashina, Anna (2014) Protein arginylation, a global biological regulator that targets actin cytoskeleton and the muscle. Anat Rec (Hoboken) 297:1630-6
Wang, Junling; Han, Xuemei; Wong, Catherine C L et al. (2014) Arginyltransferase ATE1 catalyzes midchain arginylation of proteins at side chain carboxylates in vivo. Chem Biol 21:331-7

Showing the most recent 10 out of 11 publications