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Rotavirus Genome Replication and Virion Assembly

Award Number: R01AI116815
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: 01/14/2022
PERIOD OF PERFORMANCE END DATE: 01/31/2027

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Rotavirus Genome Replication and Virion Assembly - PROJECT SUMMARY/ABSTRACT (DESCRIPTION): RNA viruses can be devastating human pathogens that impart large medical and economic burdens to society (e.g., SARS-CoV-2, influenza A virus, Ebola virus, rotavirus, etc.). While these viruses can differ quite dramatically in their pathogenesis, they share a common replication feature—they must synthesize new RNA molecules from RNA templates. Because host cell enzymes lack this activity, RNA viruses encode a specialized RNA-dependent RNA polymerase (RdRp). Viral RdRps are structurally- and functionally-conserved among diverse RNA viral families, and they directly catalyze all stages of viral transcription and genome replication. However, these enzymes rarely function alone in the context of infected cells. Instead, the viral RdRps are tightly regulated by other proteins in multi-subunit transcriptase/replicase complexes so as to maximize the type and timing of viral RNA synthesis. The overall objective of this application is to gain mechanistic insight into RdRp regulation for rotavirus—an 11-segmented, double-stranded (ds) RNA virus that causes life-threatening diarrhea in young children. The rotavirus VP1 RdRp functions only when bound beneath the icosahedral VP2 core shell layer of intact, or partially intact, particles. Engagement of VP1 by VP2 during early particle assembly activates the RdRp so that it functions as a replicase, converting packaged, single-stranded positive sense (+) RNA templates into dsRNA genome segments. Early assembly intermediates then morph into double-layered particles, wherein the VP2-bound, VP1 RdRp switches to a transcriptase activity, synthesizing +RNAs using dsRNA templates. Still, major gaps in knowledge remain about the structure of the VP1 RdRp as a replicase during dsRNA synthesis and its regulation by the VP2 core shell protein. Here, well-established in vitro biochemical and genetic techniques are combined with state-of-the-art structural approaches to close these gaps in knowledge and inform a deep understanding of rotavirus RdRp regulation. AIM 1 will elucidate VP2 core shell determinants critical for VP1 replicase activity, and AIM 2 will determine the first-ever in situ 3D atomic structures of VP1 as a replicase in both ice (using cryo-EM) and liquid (using a microfluidics system). The work outlined in this application is significant, as it is expected to reveal detailed structure-function information about the rotavirus RdRp that could be applied to rational antiviral drug design.


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 = $219,953 )
2025	2025	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	000	10	4/17/2025	NON-COMPETING CONTINUATION	$219,953
														Subtotal = $219,953

Issue Date FY: 2024 ( Subtotal = $458,225 )
2024	2024	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	001	9	9/4/2024	NON-COMPETING CONTINUATION	$45,822
2024	2024	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	000	9	1/19/2024	NON-COMPETING CONTINUATION	$412,403
														Subtotal = $458,225

Issue Date FY: 2023 ( Subtotal = $458,225 )
2023	2023	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	000	8	1/25/2023	NON-COMPETING CONTINUATION	$458,225
														Subtotal = $458,225

Issue Date FY: 2022 ( Subtotal = $463,005 )
2022	2022	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	000	7	2/18/2022	COMPETING CONTINUATION	$463,005
														Subtotal = $463,005

Issue Date FY: 2021 ( Subtotal = -$199 )
2021	2019	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	000	6	1/14/2021	NON-COMPETING CONTINUATION	-$199
														Subtotal = -$199

Issue Date FY: 2019 ( Subtotal = $321,485 )
2019	2019	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	000	6	12/10/2018	NON-COMPETING CONTINUATION	$321,485
														Subtotal = $321,485

Issue Date FY: 2018 ( Subtotal = $321,589 )
2018	2018	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	002	5	12/22/2017	NON-COMPETING CONTINUATION	$321,589
2018	2017	VIRGINIA POLYTECHNIC INSTITUTE & STATE UNIVERSITY	300 TURNER ST NW STE 4200	BLACKSBURG	VA	24061	MONTGOMERY	USA	Allergy and Infectious Diseases Research	000	3	11/14/2017	NON-COMPETING CONTINUATION	-$47,416
2018	2017	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	001	4	11/14/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$47,416
														Subtotal = $321,589

Issue Date FY: 2017 ( Subtotal = $398,777 )
2017	2017	VIRGINIA POLYTECHNIC INSTITUTE & STATE UNIVERSITY	300 TURNER ST NW STE 4200	BLACKSBURG	VA	24061	MONTGOMERY	USA	Allergy and Infectious Diseases Research	003	3	8/18/2017	NON-COMPETING CONTINUATION	-$3,292
2017	2017	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	002	4	8/1/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$154,381
2017	2017	VIRGINIA POLYTECHNIC INSTITUTE & STATE UNIVERSITY	300 TURNER ST NW, SUITE 4200	BLACKSBURG	VA	24061	MONTGOMERY	USA	Allergy and Infectious Diseases Research	001	3	8/1/2017	NON-COMPETING CONTINUATION	-$154,381
2017	2017	VIRGINIA POLYTECHNIC INSTITUTE & STATE UNIVERSITY	300 TURNER ST NW, SUITE 4200	BLACKSBURG	VA	24061	MONTGOMERY	USA	Allergy and Infectious Diseases Research	000	3	12/23/2016	NON-COMPETING CONTINUATION	$398,777
2017	2017	WAKE FOREST UNIVERSITY	1834 WAKE FOREST RD	WINSTON SALEM	NC	27109	FORSYTH	USA	Allergy and Infectious Diseases Research	004	4	8/18/2017	CHANGE OF GRANTEE / TRAINING INSTITUTION / AWARDING INSTITUTION	$3,292
														Subtotal = $398,777

Issue Date FY: 2016 ( Subtotal = $396,317 )
2016	2016	VIRGINIA POLYTECHNIC INSTITUTE & STATE UNIVERSITY	460 TURNER STREET, SUITE 306	BLACKSBURG	VA	24060	MONTGOMERY	USA	Allergy and Infectious Diseases Research	000	2	12/21/2015	NON-COMPETING CONTINUATION	$396,317
														Subtotal = $396,317

Issue Date FY: 2015 ( Subtotal = $395,287 )
2015	2015	VIRGINIA POLYTECHNIC INSTITUTE & STATE UNIVERSITY	460 TURNER STREET, SUITE 306	BLACKSBURG	VA	24060	MONTGOMERY	USA	Allergy and Infectious Diseases Research	000	1	1/14/2015	NEW	$395,287
														Subtotal = $395,287

Grand Total All Awards = $3,432,664

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Rotavirus Genome Replication and Virion Assembly

Award Number: R01AI116815

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: 01/14/2022

PERIOD OF PERFORMANCE END DATE: 01/31/2027

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