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Time-resolved Cryo-EM of Short-lived States in Eukaryotic Translation

Award Number: R01GM055440
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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Project Summary/Abstract The overall objective of this research project is the elucidation of the process of translation (i.e., translation from genetic information into protein) in eukaryotes, including mammals. Although X-ray structures exist for a number of eukaryotic ribosomes, the structural basis of this process has been mainly investigated by cryo-electron microscopy (cryo-EM). However, many short-lived states, with a life time of less than a second, cannot be imaged by standard cryo-EM (characterized by pipetting/blotting of sample), leaving large gaps in our understanding of these basic processes of life and in the knowledge base important for molecular medicine. In the approach of time-resolved cryo-EM adopted in this lab, reactions are started in a microfluidic chip (silicon- or plastic-based) by mixing two components, letting them react for a defined time (10 to 1000 ms) determined by flow rate and length of reaction channel, and the reaction product is sprayed on the EM grid immediately before the latter is plunged into the cryogen (liquid ethane, on the temperature of liquid nitrogen). In the current renewal period of this grant, ending on March 31, 2019, this technique has been greatly improved, and applied to three processes important in bacterial translation: initiation, termination, and ribosome recycling. All three applications have been successful, resulting in the capture of a short-lived state at close-to-atomic resolution and leading to papers either published or under consideration. For the current renewal, exploration of short-lived states in eukaryotic translation will have even greater relevance for human health. In collaborations with leading experts in eukaryotic translation, the Frank Lab team will apply time-resolved cryo-EM to the elucidation of processes during eukaryotic translation initiation, termination, and recycling. The aim to determine atomic structures for short-lived states that are impossible to capture by standard cryo-EM. Using these structures along with those obtained from conventional cryo-EM and X-ray crystallography, the time courses and pathways taken in the respective processes can be modeled for the first time, based on solid experiments. This new knowledge will advance strategies for combatting many diseases that implicate dysfunctions of eukaryotic translation.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2022 ( Subtotal = $0 )
2022	2020	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK, THE	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	000	23	11/6/2021	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2021 ( Subtotal = $0 )
2021	2020	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK, THE	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	000	23	1/15/2021	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2020 ( Subtotal = $346,241 )
2020	2020	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	000	23	5/19/2020	NON-COMPETING CONTINUATION	$346,241
														Subtotal = $346,241

Issue Date FY: 2019 ( Subtotal = $361,023 )
2019	2019	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	001	22	9/11/2019	COMPETING CONTINUATION	$361,436
2019	2018	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	000	21	5/23/2019	NON-COMPETING CONTINUATION	-$413
														Subtotal = $361,023

Issue Date FY: 2018 ( Subtotal = $303,913 )
2018	2018	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	000	21	3/16/2018	NON-COMPETING CONTINUATION	$273,521
2018	2018	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	001	21	5/7/2018	NON-COMPETING CONTINUATION	$30,392
														Subtotal = $303,913

Issue Date FY: 2017 ( Subtotal = $300,438 )
2017	2017	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	001	20	8/14/2017	NON-COMPETING CONTINUATION	$30,044
2017	2017	TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK	630 W 168TH ST FL 4	NEW YORK	NY	10032	NEW YORK	USA	Biomedical Research and Research Training	000	20	3/28/2017	NON-COMPETING CONTINUATION	$270,394
														Subtotal = $300,438

Issue Date FY: 2016 ( Subtotal = $296,992 )
2016	2016	COLUMBIA UNIVERSITY	615 WEST 131ST ST, 4TH FLOOR	NEW YORK	NY	10027	NEW YORK	USA	Biomedical Research and Research Training	000	19	4/4/2016	NON-COMPETING CONTINUATION	$296,992
														Subtotal = $296,992

Issue Date FY: 2015 ( Subtotal = $297,817 )
2015	2015	COLUMBIA UNIVERSITY	615 WEST 131ST ST, 4TH FLOOR	NEW YORK	NY	10027	NEW YORK	USA	Biomedical Research and Research Training	001	18	3/13/2015	COMPETING CONTINUATION	$297,817
2015	2013	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW LIBRARY	NEW YORK-NEW YORK	NY	10027		USA	Biomedical Research and Research Training	000	17	3/10/2015	NON-COMPETING CONTINUATION	$0
														Subtotal = $297,817

Issue Date FY: 2013 ( Subtotal = $304,919 )
2013	2013	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW Memorial LIBRARY	NEW YORK-NEW YORK	NY	10027		USA	Biomedical Research and Research Training	000	17	6/13/2013	NON-COMPETING CONTINUATION	$304,919
														Subtotal = $304,919

Issue Date FY: 2012 ( Subtotal = $276,001 )
2012	2012	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW Memorial LIBRARY	NEW YORK-NEW YORK	NY	10027		USA	Biomedical Research and Research Training	000	16	6/29/2012	NON-COMPETING CONTINUATION	$276,001
														Subtotal = $276,001

Issue Date FY: 2011 ( Subtotal = $275,642 )
2011	2011	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW Memorial LIBRARY	NEW YORK-NEW YORK	NY	10027		USA	Biomedical Research and Research Training	000	15	7/13/2011	NON-COMPETING CONTINUATION	$275,642
														Subtotal = $275,642

Issue Date FY: 2010 ( Subtotal = $322,686 )
2010	2010	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW Memorial LIBRARY	NEW YORK	NY	10027	NEW YORK	USA	Biomedical Research and Research Training	000	14	8/30/2010	COMPETING CONTINUATION	$322,686
2010	2009	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW LIBRARY	NEW YORK-NEW YORK	NY	10027		USA	Biomedical Research and Research Training	001	13	8/30/2010	NON-COMPETING CONTINUATION	$0
														Subtotal = $322,686

Issue Date FY: 2009 ( Subtotal = $327,681 )
2009	2009	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW Memorial LIBRARY	NEW YORK	NY	10027	NEW YORK	USA	Biomedical Research and Research Training	002	13	3/24/2009	NON-COMPETING CONTINUATION	$272,390
2009	2008	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Biomedical Research and Research Training	000	11	11/25/2008	NON-COMPETING CONTINUATION	$61,358
2009	2008	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW Memorial LIBRARY	NEW YORK	NY	10027	NEW YORK	USA	Biomedical Research and Research Training	001	12	11/25/2008	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	-$6,067
														Subtotal = $327,681

Issue Date FY: 2008 ( Subtotal = $249,228 )
2008	2008	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Biomedical Research and Research Training	000	11	3/21/2008	NON-COMPETING CONTINUATION	$95,385
2008	2008	COLUMBIA UNIVERSITY, SCHOOL OF ARTS & SCIENCES	106 LOW LIBRARY	NEW YORK-NEW YORK	NY	10027		USA	Biomedical Research and Research Training	000	12	8/27/2008	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$153,843
														Subtotal = $249,228

Issue Date FY: 2007 ( Subtotal = $256,369 )
2007	2007	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Biomedical Research and Research Training	000	10	3/19/2007	NON-COMPETING CONTINUATION	$256,369
														Subtotal = $256,369

Issue Date FY: 2006 ( Subtotal = $258,075 )
2006	2006	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Biomedical Research and Research Training	000	9	3/23/2006	COMPETING CONTINUATION	$258,075
														Subtotal = $258,075

Issue Date FY: 2004 ( Subtotal = $184,159 )
2004	2004	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Biomedical Research and Research Training	000	8	3/11/2004	NON-COMPETING CONTINUATION	$184,159
														Subtotal = $184,159

Issue Date FY: 2003 ( Subtotal = $180,077 )
2003	2003	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Cell Biology and Biophysics Research	000	7		NON-COMPETING CONTINUATION	$180,077
														Subtotal = $180,077

Issue Date FY: 2002 ( Subtotal = $176,067 )
2002	2002	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Cell Biology and Biophysics Research	000	6	3/28/2002	NON-COMPETING CONTINUATION	$176,067
														Subtotal = $176,067

Issue Date FY: 2001 ( Subtotal = $171,965 )
2001	2001	HEALTH RESEARCH, INC - ALBANY DIVISION	1 UNIVERSITY PLACE	RENSSELAER	NY	12144	RENSSELAER	USA	Cell Biology and Biophysics Research	000	5	3/9/2001	COMPETING CONTINUATION	$171,965
														Subtotal = $171,965

Issue Date FY: 2000 ( Subtotal = $181,053 )
2000	2000	HEALTH RESEARCH, INC.	RIVERVIEW CENTER	MENANDS	NY	12204	ALBANY	USA	Cell Biology and Biophysics Research	000	4	12/24/1999	NON-COMPETING CONTINUATION	$181,053
														Subtotal = $181,053

Grand Total All Awards = $5,417,001

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Time-resolved Cryo-EM of Short-lived States in Eukaryotic Translation

Award Number: R01GM055440

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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