TITLE CHANGE: FOLLOWING SINGLE MESSENGER RNA MOLECULES FROM BIRTH TO DEATH IN YEAST Over the last funding period, we have developed real-time single molecule technology that provides unparalled insight into fundamental processes of gene expression: transcription, nuclear pore export, mRNA localization and decay. The methodology permits the detection and description of kinetic events occurring at a single gene, or with a single mRNA. It has become apparent that this technology can be further extended;for instance into additional areas of mRNA metabolism or into a finer temporal and spatial regulation. This is made possible by the development of two stem-loop aptamers derived from the phages MS2 and PP7 that can be inserted into any mRNA of interest and bound to two differently colored coat proteins. These different colors can be used to mark RNA intra- or intermolecular. For instance the elongation rates of individual polymerases can be measured using these two markers, the orientation of passage of mRNAs through the nuclear pore, the translational frequency of a single mRNA, or the moment it decays. Because this single molecule technology is done in single cells, it will provide us with range of variance among cells, providing an understanding of the elasticity and tolerances in the mechanism. Because of our focus has now changed to single molecule analysis, we have changed the title to: FOLLOWING SINGLE MESSENGER RNA MOLECULES FROM BIRTH TO DEATH to reflect this change in emphasis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057071-14
Application #
8391685
Study Section
Molecular Genetics A Study Section (MGA)
Program Officer
Ainsztein, Alexandra M
Project Start
1998-01-01
Project End
2015-11-30
Budget Start
2012-12-01
Budget End
2013-11-30
Support Year
14
Fiscal Year
2013
Total Cost
$309,398
Indirect Cost
$124,130
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
Brickner, Donna Garvey; Sood, Varun; Tutucci, Evelina et al. (2016) Subnuclear positioning and interchromosomal clustering of the GAL1-10 locus are controlled by separable, interdependent mechanisms. Mol Biol Cell 27:2980-93
Vera, Maria; Biswas, Jeetayu; Senecal, Adrien et al. (2016) Single-Cell and Single-Molecule Analysis of Gene Expression Regulation. Annu Rev Genet 50:267-291
Smith, Carlas; Lari, Azra; Derrer, Carina Patrizia et al. (2015) In vivo single-particle imaging of nuclear mRNA export in budding yeast demonstrates an essential role for Mex67p. J Cell Biol 211:1121-30
Smith, Carlas S; Preibisch, Stephan; Joseph, Aviva et al. (2015) Nuclear accessibility of β-actin mRNA is measured by 3D single-molecule real-time tracking. J Cell Biol 209:609-19
Buxbaum, Adina R; Haimovich, Gal; Singer, Robert H (2015) In the right place at the right time: visualizing and understanding mRNA localization. Nat Rev Mol Cell Biol 16:95-109
Halstead, James M; Lionnet, Timothée; Wilbertz, Johannes H et al. (2015) Translation. An RNA biosensor for imaging the first round of translation from single cells to living animals. Science 347:1367-671
Campbell, Philip D; Chao, Jeffrey A; Singer, Robert H et al. (2015) Dynamic visualization of transcription and RNA subcellular localization in zebrafish. Development 142:1368-74
Grimm, Jonathan B; English, Brian P; Chen, Jiji et al. (2015) A general method to improve fluorophores for live-cell and single-molecule microscopy. Nat Methods 12:244-50, 3 p following 250
Hocine, Sami; Vera, Maria; Zenklusen, Daniel et al. (2015) Promoter-Autonomous Functioning in a Controlled Environment using Single Molecule FISH. Sci Rep 5:9934
Wu, Bin; Miskolci, Veronika; Sato, Hanae et al. (2015) Synonymous modification results in high-fidelity gene expression of repetitive protein and nucleotide sequences. Genes Dev 29:876-86

Showing the most recent 10 out of 51 publications