Mason-Pfizer monkey virus (M-PMV), is the prototype exogenous D-type retrovirus. A striking aspect of M-PMV replication is the intracytoplasmic assembly of the capsid shell. These preassembled procapsids then migrate to the plasma membrane where they associate with the glycoprotein- containing lipid bilayer. The major goal of this proposal is to obtain a better understanding of the molecular mechanisms and structural aspects involved in the process of assembling an infectious (D-type) retrovirus particle. The three major aims of this proposal are: To define using genetic and biochemical approaches structural elements in Gag that mediate intracellular transport capsid assembly and plasma membrane association. We will examine the structural basis for intracellular targeting mediated by the cytoplasmic targeting/retention signal, determine the role of matrix protein charged amino acids in membrane association and intracellular targeting, and examine functional aspects of the internal scaffold domain. Furthermore, the ability to assemble capsids in an in vitro translation system allows a direct analysis of regions involved in assembly itself rather than those that might be important for intracellular transport. To identify and define the role of host cell components in the viral assembly/intracellular transport process. We plan to utilize a combination of approaches, including yeast two hybrid systems, gene chip technology, and antibody co-precipitation to identify and characterize novel host-cell components and cellular chaperone proteins involved in the M-PMV capsid assembly/transport process. To determine the role of Env (cytoplasmic domain)/capsid interactions in the process of virus assembly. We plan to define, both structurally and functionally, the role of the cytoplasmic domain of TM in glycoprotein incorporation into M-PMV virions and examine how it regulates the fusion potential of Env. The approaches described above will extend our understanding of the molecular events involved in retrovirus assembly and will pave the way for rational approaches to develop therapeutics that can interfere with these key events in the virus life cycle.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA027834-21
Application #
6362518
Study Section
Virology Study Section (VR)
Program Officer
Read-Connole, Elizabeth Lee
Project Start
1980-09-01
Project End
2005-02-28
Budget Start
2001-03-15
Budget End
2002-02-28
Support Year
21
Fiscal Year
2001
Total Cost
$322,875
Indirect Cost
Name
University of Alabama Birmingham
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Wen, Xiaoyun; Ding, Lingmei; Wang, Jaang-Jiun et al. (2014) ROCK1 and LIM kinase modulate retrovirus particle release and cell-cell transmission events. J Virol 88:6906-21
Wen, Xiaoyun; Ding, Lingmei; Hunter, Eric et al. (2014) An siRNA screen of membrane trafficking genes highlights pathways common to HIV-1 and M-PMV virus assembly and release. PLoS One 9:e106151
Pereira, Lara E; Clark, Jasmine; Grznarova, Petra et al. (2014) Direct evidence for intracellular anterograde co-transport of M-PMV Gag and Env on microtubules. Virology 449:109-19
Clark, Jasmine; Grznarova, Petra; Stansell, Elizabeth et al. (2013) A Mason-Pfizer Monkey virus Gag-GFP fusion vector allows visualization of capsid transport in live cells and demonstrates a role for microtubules. PLoS One 8:e83863
Diehl, William E; Johnson, Welkin E; Hunter, Eric (2013) Elevated rate of fixation of endogenous retroviral elements in Haplorhini TRIM5 and TRIM22 genomic sequences: impact on transcriptional regulation. PLoS One 8:e58532
Prchal, Jan; Srb, Pavel; Hunter, Eric et al. (2012) The structure of myristoylated Mason-Pfizer monkey virus matrix protein and the role of phosphatidylinositol-(4,5)-bisphosphate in its membrane binding. J Mol Biol 423:427-38
Hadravova, Romana; de Marco, Alex; Ulbrich, Pavel et al. (2012) In vitro assembly of virus-like particles of a gammaretrovirus, the murine leukemia virus XMRV. J Virol 86:1297-306
Srb, Pavel; Vlach, Jiri; Prchal, Jan et al. (2011) Oligomerization of a retroviral matrix protein is facilitated by backbone flexibility on nanosecond time scale. J Phys Chem B 115:2634-44
Vorackova, Irena; Suchanova, Sarka; Ulbrich, Pavel et al. (2011) Purification of proteins containing zinc finger domains using immobilized metal ion affinity chromatography. Protein Expr Purif 79:88-95
Prchal, Jan; Junkova, Petra; Strmiskova, Miroslava et al. (2011) Expression and purification of myristoylated matrix protein of Mason-Pfizer monkey virus for NMR and MS measurements. Protein Expr Purif 79:122-7

Showing the most recent 10 out of 62 publications