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
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Vera, Maria; Pani, Bibhusita; Griffiths, Lowri A et al. (2014) The translation elongation factor eEF1A1 couples transcription to translation during heat shock response. Elife 3:e03164
Hocine, Sami; Raymond, Pascal; Zenklusen, Daniel et al. (2013) Single-molecule analysis of gene expression using two-color RNA labeling in live yeast. Nat Methods 10:119-21
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Haimovich, Gal; Choder, Mordechai; Singer, Robert H et al. (2013) The fate of the messenger is pre-determined: a new model for regulation of gene expression. Biochim Biophys Acta 1829:643-53
Lionnet, Timothee; Singer, Robert H (2012) Transcription goes digital. EMBO Rep 13:313-21
Wu, Bin; Piatkevich, Kiryl D; Lionnet, Timothee et al. (2011) Modern fluorescent proteins and imaging technologies to study gene expression, nuclear localization, and dynamics. Curr Opin Cell Biol 23:310-7
Larson, Daniel R; Zenklusen, Daniel; Wu, Bin et al. (2011) Real-time observation of transcription initiation and elongation on an endogenous yeast gene. Science 332:475-8

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