Cell migration drives such key biological events as tissue morphogenesis, immune response, and cancer metastases. Our recent data surprisingly show that the formation and function of the cell leading edge during migration critically depends on arginylation, a relatively unexplored posttranslational modification. Moreover, in cell-based phenotype rescue experiments, we have shown that experimentally arginylated beta actin can largely restore cell leading edge function in mouse embryonic fibroblasts lacking the arginylation enzyme ATE1. In our ongoing studies to determine how N-terminal arginylation of beta actin contributes to lamellipodia formation and directed cell migration, we have made the novel observations that a prominent subset of arginylated beta actin is targeted to the cell leading edge during migration. Moreover, our data demonstrate that beta actin arginylation is selectively regulated by its mRNA sequence rather than its protein structure. In support, gamma actin, which is 99% identical to beta actin at the amino acid level, differs by 13% in its mRNA sequence. We have shown that this difference is directly responsible for faster translation rate of beta actin and leads to its selective arginylation through a novel mechanism coupled to protein ubiquitination. It is also known that zipcode-mediated beta actin mRNA targeting regulates its leading edge localization and, like arginylation, is essential for directional cell migration. We hypothesize that mRNA-mediated regulation of N-terminal arginylation of beta actin uniquely regulates actin function during cell migration by facilitating actin polymerization at the cell leading edge. In this proposal, we will test this hypothesis through three specific aims that will: (1) test the hypothesis that beta actin arginylation facilitates actin polymerization at the cell leading edge; (2) test the hypothesis that beta actin function is uniquely regulated by coding and noncoding regions of its mRNA; and (3) test the hypothesis that this regulation is coupled to modulation of intracellular arginylation activity during cell migration. These experiments will address a novel regulatory mechanism controlling cell polarization and motility through modulating actin's properties and mRNA structure and will ultimately enable a new level of targeted functional studies of cell migration during essential physiological events.

Public Health Relevance

Abnormalities in actin cytoskeleton result in defects in cell migration and adhesion that lead to severe developmental defects and contribute to numerous 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 novel regulation of the actin cytoskeleton by posttranslational arginylation, with the goal of enabling the development of new therapeutics that regulate aberrant cell migration and adhesion in these disease states.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM108744-04
Application #
9215681
Study Section
Nuclear and Cytoplasmic Structure/Function and Dynamics Study Section (NCSD)
Program Officer
Nie, Zhongzhen
Project Start
2014-04-01
Project End
2018-02-28
Budget Start
2017-03-01
Budget End
2018-02-28
Support Year
4
Fiscal Year
2017
Total Cost
$273,600
Indirect Cost
$102,600
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
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
Cornachione, Anabelle S; Leite, Felipe S; Wang, Junling et al. (2014) Arginylation of myosin heavy chain regulates skeletal muscle strength. Cell Rep 8:470-6