The purpose of this proposal is to understand the role of zipcode-binding protein ZBP1 in the mechanism of mRNA localization. Over the last project period, we have identified and characterized the role of this protein in localizing ?-actin mRNA to the periphery of cells where it is involved in synthesizing proteins important for neuronal growth and cell motility. This protein binds to the """"""""zipcode"""""""" of mRNA and transduces nucleic acid sequences into cellular spatial information. Surprisingly, this protein is at a central point in the cellular basis of disease, we have found it to be a metastasis suppressor and play a role in neuronal pathology and even in behavior. We hypothesize that ZBP1 is carrying a group of mRNAs to focal adhesion complexes or sites of cell-cell contact and this regulates intercellular or cell-substrate interactions. Our experimental focus in this work is to identify the RNA and protein binding partners of ZBP1;in particular using TIRF. We will then characterize its crystal structure and determine how it interacts with specific proteins to move mRNAs and allow their expression. Finally, we will visualize these processes in living cells from mice carrying a modified ?-actin mRNA and in a knock-out of ZBP1.

Public Health Relevance

We have discovered a protein that controls how and where other proteins are made in the cell. We have found that this protein is implicated in suppressing cancer metastasis and maintaining normal nerve function. We are investigating how this protein works to localize protein synthesis in specific regions of the cell by using a super-sensitive microscope to watch the process occur in living cells and tissues from genetically engineered mice.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM084364-18S1
Application #
8536993
Study Section
Nuclear Dynamics and Transport (NDT)
Program Officer
Ainsztein, Alexandra M
Project Start
1992-03-03
Project End
2013-07-31
Budget Start
2011-08-01
Budget End
2013-07-31
Support Year
18
Fiscal Year
2012
Total Cost
$384,641
Indirect Cost
$154,317
Name
Albert Einstein College of Medicine
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Yoon, Young J; Wu, Bin; Buxbaum, Adina R et al. (2016) Glutamate-induced RNA localization and translation in neurons. Proc Natl Acad Sci U S A 113:E6877-E6886
Wang, Guangli; Huang, Zhenqiang; Liu, Xin et al. (2016) IMP1 suppresses breast tumor growth and metastasis through the regulation of its target mRNAs. Oncotarget 7:15690-702
Song, Tingting; Zheng, Yi; Wang, Yarong et al. (2015) Specific interaction of KIF11 with ZBP1 regulates the transport of β-actin mRNA and cell motility. J Cell Sci 128:1001-10
Buxbaum, Adina R; Wu, Bin; Singer, Robert H (2014) Single β-actin mRNA detection in neurons reveals a mechanism for regulating its translatability. Science 343:419-22
Park, Hye Yoon; Lim, Hyungsik; Yoon, Young J et al. (2014) Visualization of dynamics of single endogenous mRNA labeled in live mouse. Science 343:422-4
Wu, Bin; Chen, Jiahao; Singer, Robert H (2014) Background free imaging of single mRNAs in live cells using split fluorescent proteins. Sci Rep 4:3615
Coulon, Antoine; Chow, Carson C; Singer, Robert H et al. (2013) Eukaryotic transcriptional dynamics: from single molecules to cell populations. Nat Rev Genet 14:572-84
Trcek, Tatjana; Sato, Hanae; Singer, Robert H et al. (2013) Temporal and spatial characterization of nonsense-mediated mRNA decay. Genes Dev 27:541-51
Todorova, Nevena; Makarucha, Adam J; Hine, Nicholas D M et al. (2013) Dimensionality of carbon nanomaterials determines the binding and dynamics of amyloidogenic peptides: multiscale theoretical simulations. PLoS Comput Biol 9:e1003360
Wu, Bin; Singer, Robert H; Mueller, Joachim D (2013) Time-integrated fluorescence cumulant analysis and its application in living cells. Methods Enzymol 518:99-119

Showing the most recent 10 out of 33 publications