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ASSEMBLY OF ENVELOPED VIRUSES

Award Number: R01AI015892
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: 05/01/1979
PERIOD OF PERFORMANCE END DATE: 06/30/2014

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ASSEMBLY OF ENVELOPED VIRUSES - Project Abstract The goal of this project is to determine the mechanism of assembly of the envelope of vesicular stomatitis virus (VSV), the prototype rhabdovirus, which has been widely used to study virus assembly. Like many enveloped viruses, VSV has a matrix (M) protein, which serves as a bridge between the nucleocapsid and the virus envelope. The mechanism of how nucleocapsids and M proteins are assembled at the cytoplasmic surface of host membranes to form virus budding sites is a central question in virus assembly. According to virology textbooks, assembly at the plasma membrane is mediated through a transmembrane network of protein- protein interactions of M protein with nucleocapsids and the envelope glycoprotein (G protein). However, most of the available evidence in the literature argues against this textbook model of virus assembly. The research in this project has developed novel methods of electron microscopy analysis and new biophysical and biochemical tools, which have provided evidence for a new model for assembly of VSV envelopes, as follows: A key step is movement of viral nucleocapsids to membrane-proximal regions of the cytoplasm, where they associate with membrane microdomains that are enriched not in M protein, but rather in G protein. 2) These membrane microdomains serve as sites where association of M protein with nucleocapsids occurs. 3) Importantly, one or more of these steps is regulated by host factors that affect virus assembly. This model will be tested by the following aims: Aim 1 is to test the hypothesis that a key step in the initiation of virus assembly is association of viral nucleocapsids with membrane microdomains that are enriched in G protein. These experiments will use new methods of confocal and electron microscopy analysis to study previously described virus mutants, in which assembly of nucleocapsids into virions is defective. The model will also be tested by complementary biochemical approaches and a modification of the "microdomain proteomics" approach to identify host proteins associated with sites of virus assembly. In Aim 2 the mechanism of assembly of M protein into nucleocapsid-M protein (NCM) complexes will be tested. These experiments will use novel stopped-flow light scattering approaches capable of measuring the rapid rates of these protein-protein interactions. This Aim will also analyze the ability of mutant M proteins to be recruited into NCM complexes in vivo. These experiments will involve complementation of temperature-sensitive M protein mutants, which have been shown previously to be defective in assembly of M protein into virions. These experiments not only provide an in vivo assay for assembly of NCM complexes, but also have the potential to provide important new tools for studying the initiating steps in virus assembly.


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: 2013 ( Subtotal = $340,880 )
2013	2013	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	32	6/17/2013	NON-COMPETING CONTINUATION	$340,880
														Subtotal = $340,880

Issue Date FY: 2012 ( Subtotal = $362,637 )
2012	2012	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	31	6/13/2012	NON-COMPETING CONTINUATION	$362,637
														Subtotal = $362,637

Issue Date FY: 2011 ( Subtotal = $362,637 )
2011	2011	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	30	6/14/2011	NON-COMPETING CONTINUATION	$362,637
														Subtotal = $362,637

Issue Date FY: 2010 ( Subtotal = $366,300 )
2010	2010	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	29	6/3/2010	NON-COMPETING CONTINUATION	$366,300
														Subtotal = $366,300

Issue Date FY: 2009 ( Subtotal = $370,000 )
2009	2009	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	001	28	6/8/2009	COMPETING CONTINUATION	$370,000
2009	2008	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	27	6/8/2009	NON-COMPETING CONTINUATION	$0
														Subtotal = $370,000

Issue Date FY: 2008 ( Subtotal = $266,957 )
2008	2008	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	27	12/13/2007	NON-COMPETING CONTINUATION	$266,957
														Subtotal = $266,957

Issue Date FY: 2007 ( Subtotal = $272,128 )
2007	2007	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	26	12/19/2006	NON-COMPETING CONTINUATION	$272,128
														Subtotal = $272,128

Issue Date FY: 2006 ( Subtotal = $280,256 )
2006	2006	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Allergy and Infectious Diseases Research	000	25	12/12/2005	NON-COMPETING CONTINUATION	$280,256
														Subtotal = $280,256

Issue Date FY: 2005 ( Subtotal = $287,000 )
2005	2005	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Microbiology and Infectious Diseases Research	000	24	1/24/2005	NON-COMPETING CONTINUATION	$287,000
														Subtotal = $287,000

Issue Date FY: 2004 ( Subtotal = $287,500 )
2004	2004	WAKE FOREST UNIVERSITY HEALTH SCIENCES	MEDICAL CENTER BOULEVARD	WINSTON SALEM	NC	27157	FORSYTH	USA	Microbiology and Infectious Diseases Research	000	23	12/22/2003	COMPETING CONTINUATION	$287,500
														Subtotal = $287,500

Issue Date FY: 2002 ( Subtotal = $262,763 )
2002	2002	WAKE FOREST UNIVERSITY SCHOOL OF MEDICINE	MEDICAL CENTER BOULEVARD	WINSTON-SALEM	NC	27157		USA	Microbiology and Infectious Diseases Research	000	22	1/23/2002	NON-COMPETING CONTINUATION	$262,763
														Subtotal = $262,763

Issue Date FY: 2001 ( Subtotal = $255,110 )
2001	2001	WAKE FOREST UNIVERSITY SCHOOL OF MEDICINE	MEDICAL CENTER BOULEVARD	WINSTON-SALEM	NC	27157		USA	Microbiology and Infectious Diseases Research	000	21	1/5/2001	NON-COMPETING CONTINUATION	$255,110
														Subtotal = $255,110

Issue Date FY: 2000 ( Subtotal = $247,679 )
2000	2000	WAKE FOREST UNIVERSITY SCHOOL OF MEDICINE	MEDICAL CENTER BOULEVARD	WINSTON-SALEM	NC	27157		USA	Microbiology and Infectious Diseases Research	000	20	1/24/2000	NON-COMPETING CONTINUATION	$247,679
														Subtotal = $247,679

Issue Date FY: 1999 ( Subtotal = $240,465 )
1999	1999	WAKE FOREST UNIVERSITY SCHOOL OF MEDICINE	MEDICAL CENTER BOULEVARD	WINSTON-SALEM	NC	27157		USA	Microbiology and Infectious Diseases Research	000	19	1/25/1999	NON-COMPETING CONTINUATION	$240,465
														Subtotal = $240,465

Issue Date FY: 1998 ( Subtotal = $233,462 )
1998	1998	WAKE FOREST UNIVERSITY SCHOOL OF MEDICINE	MEDICAL CENTER BOULEVARD	WINSTON-SALEM	NC	27157		USA	Microbiology and Infectious Diseases Research	000	18	2/6/1998	COMPETING CONTINUATION	$233,462
														Subtotal = $233,462

Grand Total All Awards = $4,435,774

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ASSEMBLY OF ENVELOPED VIRUSES

Award Number: R01AI015892

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: 05/01/1979

PERIOD OF PERFORMANCE END DATE: 06/30/2014

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