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Dissection of molecular events driving alphavirus entry and assembly

Award Number: R01AI075647
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/15/1995
PERIOD OF PERFORMANCE END DATE: 11/30/2024

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Dissection of molecular events driving alphavirus entry and assembly - Alphaviruses include important and emerging human pathogens such as Chikungunya virus, Mayaro virus and the encephalitic alphaviruses. There are currently no licensed antiviral therapies or vaccines, and new approaches and information are needed to develop antiviral strategies. Alphaviruses are small enveloped plus-sense RNA viruses that infect cells via clathrin-mediated uptake and low pH-triggered fusion, and replicate in the cytoplasm. The capsid protein selectively packages the RNA genome to form the highly organized nucleocapsid. Budding through the plasma membrane envelopes this core in a membrane containing an organized lattice of the E2/E1 envelope proteins. The resulting virus particle has high specific infectivity, attesting to the specificity and high-quality control of assembly. Gaps in our knowledge include the mechanisms that promote and constrain RNA packaging and nucleocapsid formation, which occur in the presence of abundant cellular and subgenomic RNAs and high-levels of cytoplasmic capsid protein. While it is clear that alphavirus entry involves receptor-mediated endocytosis, there is little molecular information on virus-receptor interactions. This grant will address these critical steps in 3 inter-related aims: 1. Mechanism by which berberine inhibits alphavirus nucleocapsid formation. Our preliminary data identify a novel and selective effect of the small molecule berberine on alphavirus nucleocapsid formation. We will define berberine’s mechanism and use this inhibitor to analyze the biology of nucleocapsid assembly. 2. RNA-capsid protein interactions that drive selective viral RNA packaging in virus-infected cells. We have used 4-thiouridine labeling and photoactivation to specifically crosslink viral RNA with capsid protein in the cytoplasmic, cellular nucleocapsid and viral nucleocapsid pools. Our preliminary results indicate that we have mapped capsid binding sites on the viral RNA with nucleotide precision, and identified both the known packaging sequence and a set of novel sites in the genomic RNA. We will analyze these sites for sequence/structural motifs and compare across alphavirus clades. We will test the roles of these novel sites in mediating specific viral RNA packaging, nucleocapsid assembly, and packaging of reporter RNAs. 3. Molecular features of NRAMP2 as an alphavirus receptor. The multi-pass iron transporter NRAMP2 has been identified as the receptor for Sindbis virus (SINV), but the structure and function of the receptor vs. virus infection are not understood. We will determine the properties of the receptor required to promote endocytic uptake and infection. We will take advantage of recent NRAMP structural studies to test the role of iron binding, pH regulation, and transporter conformational changes in virus receptor activity, and map the SINV binding sites on the extracellular loop(s) of NRAMP2. The results of these aims will provide novel and fundamental information on the alphavirus lifecycle and the host factors that promote it.


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: 2025 ( Subtotal = -$451 )
2025	2023	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	29	4/15/2025	NON-COMPETING CONTINUATION	-$451
														Subtotal = -$451

Issue Date FY: 2023 ( Subtotal = $633,513 )
2023	2023	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	29	11/30/2022	NON-COMPETING CONTINUATION	$633,513
														Subtotal = $633,513

Issue Date FY: 2022 ( Subtotal = $633,513 )
2022	2022	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	28	11/4/2021	NON-COMPETING CONTINUATION	$633,513
														Subtotal = $633,513

Issue Date FY: 2021 ( Subtotal = $633,513 )
2021	2021	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	27	11/20/2020	NON-COMPETING CONTINUATION	$633,513
														Subtotal = $633,513

Issue Date FY: 2020 ( Subtotal = $633,513 )
2020	2020	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	26	11/27/2019	NON-COMPETING CONTINUATION	$633,513
														Subtotal = $633,513

Issue Date FY: 2019 ( Subtotal = $613,287 )
2019	2019	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	003	25	2/8/2019	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$536,410
2019	2019	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	001	24	11/20/2018	COMPETING CONTINUATION	$613,287
2019	2019	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	005	24	4/25/2019	COMPETING CONTINUATION	$134,610
2019	2019	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	002	24	2/8/2019	COMPETING CONTINUATION	-$536,410
2019	2019	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	007	25	5/9/2019	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$189,597
2019	2019	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	006	24	5/9/2019	COMPETING CONTINUATION	-$189,597
2019	2017	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	23	11/19/2018	NON-COMPETING CONTINUATION	$0
2019	2017	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	004	23	4/25/2019	NON-COMPETING CONTINUATION	-$134,610
														Subtotal = $613,287

Issue Date FY: 2017 ( Subtotal = $925,401 )
2017	2017	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	007	23	3/15/2017	NON-COMPETING CONTINUATION	$653,224
2017	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	002	21	10/25/2016	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$272,177
2017	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	005	21	10/28/2016	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$263,240
2017	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE, INC.	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	006	21	10/28/2016	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$8,937
2017	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	001	20	10/25/2016	NON-COMPETING CONTINUATION	$8,937
2017	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	004	20	10/28/2016	NON-COMPETING CONTINUATION	-$263,240
2017	2014	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	003	19	10/28/2016	NON-COMPETING CONTINUATION	-$8,937
2017	2014	ALBERT EINSTEIN COLLEGE OF MEDICINE	1300 MORRIS PARK AVE	BRONX	NY	10461	BRONX	USA	Allergy and Infectious Diseases Research	000	19	10/25/2016	NON-COMPETING CONTINUATION	-$8,937
														Subtotal = $925,401

Issue Date FY: 2016 ( Subtotal = $653,224 )
2016	2016	ALBERT EINSTEIN COLLEGE OF MEDICINE INC	111 E 210TH ST	BRONX	NY	10467	BRONX	USA	Allergy and Infectious Diseases Research	001	22	3/4/2016	NON-COMPETING CONTINUATION	$653,224
2016	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE INC	111 E 210TH ST	BRONX	NY	104672401	BRONX	USA	Allergy and Infectious Diseases Research	000	21	2/10/2016	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$0
														Subtotal = $653,224

Issue Date FY: 2015 ( Subtotal = $653,224 )
2015	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	002	20	9/14/2015	NON-COMPETING CONTINUATION	-$381,047
2015	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	20	3/12/2015	NON-COMPETING CONTINUATION	$653,224
2015	2015	ALBERT EINSTEIN COLLEGE OF MEDICINE INC	111 E 210TH ST	BRONX	NY	10467	BRONX	USA	Allergy and Infectious Diseases Research	001	21	9/14/2015	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$381,047
														Subtotal = $653,224

Issue Date FY: 2014 ( Subtotal = $653,224 )
2014	2014	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	19	3/7/2014	NON-COMPETING CONTINUATION	$653,224
														Subtotal = $653,224

Issue Date FY: 2013 ( Subtotal = $599,881 )
2013	2013	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	002	18	7/12/2013	COMPETING CONTINUATION	$25,526
2013	2013	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	001	18	3/13/2013	COMPETING CONTINUATION	$574,355
2013	2011	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	17	3/11/2013	NON-COMPETING CONTINUATION	$0
														Subtotal = $599,881

Issue Date FY: 2011 ( Subtotal = $540,778 )
2011	2011	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	17	7/29/2011	NON-COMPETING CONTINUATION	$540,778
														Subtotal = $540,778

Issue Date FY: 2010 ( Subtotal = $535,843 )
2010	2010	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	16	6/21/2010	NON-COMPETING CONTINUATION	$535,843
														Subtotal = $535,843

Issue Date FY: 2009 ( Subtotal = $586,216 )
2009	2009	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Trans-NIH Recovery Act Research Support	001	15	9/7/2009	SUPPLEMENT FOR EXPANSION	$60,584
2009	2009	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	15	7/24/2009	NON-COMPETING CONTINUATION	$525,632
														Subtotal = $586,216

Issue Date FY: 2008 ( Subtotal = $510,692 )
2008	2008	ALBERT EINSTEIN COLLEGE OF MEDICINE OF YESHIVA UNIV	1300 MORRIS PARK AVENUE	BRONIX	NY	10461		USA	Allergy and Infectious Diseases Research	000	14	8/12/2008	NON-COMPETING CONTINUATION	$510,692
														Subtotal = $510,692

Grand Total All Awards = $9,310,712

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Dissection of molecular events driving alphavirus entry and assembly

Award Number: R01AI075647

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 08/15/1995

PERIOD OF PERFORMANCE END DATE: 11/30/2024

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