Dependovirus; Biochemistry; Eukaryotic Cells; Genetics; Melanoma, Experimental; Mice, Mutant Strains; Medical Laboratory Science; Parvovirus; Erythrovirus; Oncolytic Virotherapy; Oncolytic Viruses; Mice; Bocavirus; Viral Structures; Immunomodulation
Molecular Virology: Virology laboratories
Our research efforts are directed at understanding the molecular mechanisms by which mammalian parvoviruses target particular cell types, express their genes, take over their host cells and replicate their own DNA. Eukaryotic and prokaryotic expression systems, coupled with directed mutagenesis, are currently being used to separate the various functions of the complex, multi-functional parvoviral gene products, in order to understand how the virus subverts the macromolecular metabolism of its target host cell to its own ends. We are currently applying this knowledge to the construction of vectors for transducing immunomodulatory genes into tumor cells as therapeutic strategy against cancer.
Specialized Terms: Biochemistry; Genetics; Parvoviruses; DNA replication; Gene therapy; Oncolytic virus; Vaccines; Vectors; Viral replication and vectors
Extensive Research Description
Manipulating the oncoselectivity of parvoviruses in human tumor models
Most of the rodent parvoviruses will bind to and enter human cells with high efficiency, but fail to initiate gene expression, replicate their genomes, generate progeny or spread through the culture, unless the host cell is neoplastically transformed. As a consequence, these viruses are promising candidates as oncolytic agents for cancer therapy, particularly in situations where other treatments have proven ineffective. Our current efforts are directed toward understanding, at the molecular level, why cellular changes that accompany oncogenic transformation promote viral growth, and how we can use this knowledge to further improve the efficacy of the virus in tumor eradication. Since tumorigenesis normally involves loss of genomic integrity, tumor cells carry many mutations that are secondary to those causing the transformed phenotype. To avoid studying or selecting for viral traits that represent adaptations to such “collateral” transformed cell properties, we are using host cells that have been transformed in a stepwise fashion with activated oncogenes and/or tumor suppressor knock-downs. Currently we are exploring the contribution of the viral capsid and initiating promoter to the discrimination between normal and transformed cells, using stepwise transformed human fibroblasts and melanocytes, the latter being a model for malignant melanoma. These studies are directing strategies for selecting more oncotropic versions of these critical oncoselective elements, using gene shuffling and degenerate promoter library approaches.
Induction of immunogenic cell death by oncosuppressive parvoviruses
Many of the autonomously replicating rodent parvoviruses can enter human cells, generate progeny and spread through the culture only if the host cell is neoplastically transformed, making these viruses promising candidates as oncolytic agents. Parvoviral induction of complete tumor regression has been achieved in several syngeneic transplantable tumor models in immunocompetent rodent hosts, and often results in immunization of the animal against subsequent transplantation of cells of the same tumor, even at high input numbers, suggesting that some aspect of parvovirus infection elicits a strong anti-tumor immune response. This project utilizes a mouse melanoma model system to explore whether parvovirus-induced cell death proceeds via an immunogenic, rather than tolerogenic, pathway, by examining the expression of phagocytic engulfment signals on the infected cell surface, coupled with the secretion of soluble damage-associated molecular pattern (DAMP) molecules, such as HMGB1 and Hsp72.
How parvoviruses enter their host cell and traffic to the nucleus
Parvoviruses do not have a lipid envelope, and so cannot deliver their virions into the host cell by fusing with its plasma or endosomal membranes. These viruses have developed an alternative strategy to breach their host cell's outer membrane and gain entry into the cytoplasm. We have shown that the compact, icosahedral virion of the murine parvovirus Minute Virus of Mice, MVM deploys a lipolytic enzyme, phospholipase A2 (PLA2) that is expressed at the N-terminus of the minor coat protein, VP1. This region of VP1 is normally sequestered within the viral shell, but is extruded during the entry process as a capsid-tethered domain, via an 8Å pore that extends through the prominent 5-fold cylinder. [Figure] In addition to the PLA2 domain, the extruded VP1 N-terminus also displays a number of small protein interaction domains predicted to engage both ubiquitin ligases of the NEDD4 family, involved in endocytosis and vesicle trafficking, and nuclear transport proteins of the alpha-importin family. The sequential conformational shifts within the particle that allow these transitions to occur as the virion transits its entry pathway, exposing first its VP2 N-termini, then its VP1 N-termini and ultimately its DNA are being analyzed using X-ray crystallography and asymmetric cryo-electron microscopy, in collaborations with Drs. Susan Hafenstein at Hershey Medical School and Mavis Agbandje-McKenna at the University of Florida, Gainesville. Finally, we are using reverse genetics combined with differential real-time PCR, sub-cellular fractionation and in situ imaging techniques, such as Proximity Ligation, to explore the roles of the VP1 N-terminal domain in the trans-cytosolic trafficking and nuclear import of MVM virions.
Characterizing the unique chromatin assembled during parvoviral DNA replication
During the S-phase following infection, autonomous parvoviruses inveigle host cells to replicate the linear, single-stranded viral DNA chromosome, instead of the cellular genome. Part of the virus’ replication strategy involves the elaboration of a unique form of chromatin, which ChIP analysis suggests incorporates both cellular histones and many copies of NS1, the major viral non-structural protein. This project explores the replication of an otherwise wildtype viral genome rendered artificially devoid of NS1 binding sites throughout its entire NS1 gene, capsid gene and/or 3’ untranslated region. 2D gel electrophoresis and nuclease protection assays will be used to look for stalling or pausing of replication forks through the capsid region, and to characterize packaging intermediates generated by mutant, compared to wildtype, virus.
Complementation for an essential ancillary non-structural protein function across parvovirus genera.
Mihaylov, I.S., Cotmore, S.F., & Tattersall, P. Complementation for an essential ancillary non-structural protein function across parvovirus genera. Virology. 468-470:226-237, 2014.
Parvoviral left-end hairpin ears are essential during infection for establishing a functional intranuclear transcription template and for efficient progeny genome encapsidation.
Li, L., Cotmore, S.F., & Tattersall, P. Parvoviral left-end hairpin ears are essential during infection for establishing a functional intranuclear transcription template and for efficient progeny genome encapsidation. J. Virol. 87:10501-14, 2013.
Distinct host cell fates for human malignant melanoma targeted by oncolytic rodent parvoviruses.
Vollmers, E.M., & Tattersall, P. Distinct host cell fates for human malignant melanoma targeted by oncolytic rodent parvoviruses. Virology, 446:37-48, 2013.
Parvovirus evades interferon-dependent viral control in primary mouse embryonic fibroblasts.
Mattei, L.M., Cotmore, S.F., Tattersall, P., & Iwasaki, A. Parvovirus evades interferon-dependent viral control in primary mouse embryonic fibroblasts. Virology, 442:20-7, 2013
Mutations at the base of the icosahedral five-fold cylinders of Minute Virus of Mice induce 3'-to-5' genome uncoating and critically impair entry functions.
Cotmore, S.F. & Tattersall, P. Mutations at the base of the icosahedral five-fold cylinders of Minute Virus of Mice induce 3'-to-5' genome uncoating and critically impair entry functions. J. Virol., 86: 69-80, 2012.
The parvoviral capsid controls an intracellular phase of infection essential for efficient killing of stepwise-transformed human fibroblasts.
Paglino, J. & Tattersall, P. The parvoviral capsid controls an intracellular phase of infection essential for efficient killing of stepwise-transformed human fibroblasts. Virology, 416:32-41, 2011.
Structure of a packaging-defective mutant of Minute Virus of Mice indicates that the genome is packaged via a pore at a 5-fold axis.
Plevka, P., Hafenstein, S., Li, L., D'Abramo, A. Jr., Cotmore, S.F., Rossmann, M.G., & Tattersall P. Structure of a packaging-defective mutant of Minute Virus of Mice indicates that the genome is packaged via a pore at a 5-fold axis. J. Virol., 85:4822-7, 2011.
Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM).
Ruiz, Z., Mihaylov, I.S., Cotmore, S.F., & Tattersall, P. Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM). Virology, 410:375-84, 2011.
Structures of minute virus of mice replication initiator protein N-terminal domain: Insights into DNA nicking and origin binding
Tewary, S.K., Liang, L., Lin, Z., Lynn, A., Cotmore, S.F., Tattersall, P., Zhao, H. & Tang, L. Virology 476:61-71, 2015.
The MVMp P4 promoter is a host cell-type range determinant in vivo
Meir, C., Mincberg, M., Rostovsky, I., Tal, S., Vollmers, E.M., Levi, A,. Tattersall, P. & Davis, C. Virology 506:141-51, 2017.
Cryo-EM maps reveal five-fold channel structures and their modification by gatekeeper mutations in the parvovirus minute virus of mice (MVM) capsid
Subramanian, S., Organtini ,L.J., Grossman, A., Domeier, P.P., Cifuente, J.O., Makhov, A.M., Conway, J.F., D'Abramo, A., Jr., Cotmore, S.F., Tattersall, P. & Hafenstein, S. Virology 510:216-223, 2017.
Full List of PubMed Publications
- Pittman N, Misseldine A, Geilen L, Halder S, Smith JK, Kurian J, Chipman P, Janssen M, Mckenna R, Baker TS, D'Abramo A Jr, Cotmore S, Tattersall P, Agbandje-McKenna M: Atomic Resolution Structure of the Oncolytic Parvovirus LuIII by Electron Microscopy and 3D Image Reconstruction. Viruses. 2017 Oct 30; 2017 Oct 30. PMID: 29084163
- Subramanian S, Organtini LJ, Grossman A, Domeier PP, Cifuente JO, Makhov AM, Conway JF, D'Abramo A Jr, Cotmore SF, Tattersall P, Hafenstein S: Cryo-EM maps reveal five-fold channel structures and their modification by gatekeeper mutations in the parvovirus minute virus of mice (MVM) capsid. Virology. 2017 Oct; 2017 Jul 24. PMID: 28750325
- Meir C, Mincberg M, Rostovsky I, Tal S, Vollmers EM, Levi A, Tattersall P, Davis C: The MVMp P4 promoter is a host cell-type range determinant in vivo. Virology. 2017 Jun; 2017 Apr 6. PMID: 28391161
- Nakaya Y, Stavrou S, Blouch K, Tattersall P, Ross SR: In Vivo Examination of Mouse APOBEC3- and Human APOBEC3A- and APOBEC3G-Mediated Restriction of Parvovirus and Herpesvirus Infection in Mouse Models. J Virol. 2016 Sep 1; 2016 Aug 12. PMID: 27356895
- Tewary SK, Liang L, Lin Z, Lynn A, Cotmore SF, Tattersall P, Zhao H, Tang L: Structures of minute virus of mice replication initiator protein N-terminal domain: Insights into DNA nicking and origin binding. Virology. 2015 Feb; 2014 Dec 18. PMID: 25528417
- Cotmore SF, Tattersall P: Parvoviruses: Small Does Not Mean Simple. Annu Rev Virol. 2014 Nov; 2014 Jul 9. PMID: 26958732
- Mihaylov IS, Cotmore SF, Tattersall P: Complementation for an essential ancillary non-structural protein function across parvovirus genera. Virology. 2014 Nov; 2014 Sep 6. PMID: 25194919
- Vollmers EM, D'Abramo A Jr, Cotmore SF, Tattersall P: Genome sequence of tumor virus x, a member of the genus protoparvovirus in the family parvoviridae. Genome Announc. 2014 Jul 31; 2014 Jul 31. PMID: 25081268
- Cotmore SF, Agbandje-McKenna M, Chiorini JA, Mukha DV, Pintel DJ, Qiu J, Soderlund-Venermo M, Tattersall P, Tijssen P, Gatherer D, Davison AJ: The family Parvoviridae. Arch Virol. 2014 May; 2013 Nov 9. PMID: 24212889
- Halder S, Cotmore S, Heimburg-Molinaro J, Smith DF, Cummings RD, Chen X, Trollope AJ, North SJ, Haslam SM, Dell A, Tattersall P, McKenna R, Agbandje-McKenna M: Profiling of glycan receptors for minute virus of mice in permissive cell lines towards understanding the mechanism of cell recognition. PLoS One. 2014; 2014 Jan 27. PMID: 24475195
- Vollmers EM, Tattersall P: Distinct host cell fates for human malignant melanoma targeted by oncolytic rodent parvoviruses. Virology. 2013 Nov; 2013 Aug 9. PMID: 24074565
- Mattei LM, Cotmore SF, Tattersall P, Iwasaki A: Parvovirus evades interferon-dependent viral control in primary mouse embryonic fibroblasts. Virology. 2013 Jul 20; 2013 May 12. PMID: 23676303
- Mattei LM, Cotmore SF, Li L, Tattersall P, Iwasaki A: Toll-like receptor 9 in plasmacytoid dendritic cells fails to detect parvoviruses. J Virol. 2013 Mar; 2013 Jan 9. PMID: 23302877
- Cotmore SF, Tattersall P: Parvovirus diversity and DNA damage responses. Cold Spring Harb Perspect Biol. 2013 Feb 1; 2013 Feb 1. PMID: 23293137
- Li L, Cotmore SF, Tattersall P: Maintenance of the flip sequence orientation of the ears in the parvoviral left-end hairpin is a nonessential consequence of the critical asymmetry in the hairpin stem. J Virol. 2012 Nov; 2012 Aug 29. PMID: 22933276
- Cotmore SF, Tattersall P: Mutations at the base of the icosahedral five-fold cylinders of minute virus of mice induce 3'-to-5' genome uncoating and critically impair entry functions. J Virol. 2012 Jan; 2011 Oct 19. PMID: 22013064
- Paglino J, Tattersall P: The parvoviral capsid controls an intracellular phase of infection essential for efficient killing of stepwise-transformed human fibroblasts. Virology. 2011 Jul 20; 2011 May 20. PMID: 21600623
- Ruiz Z, Mihaylov IS, Cotmore SF, Tattersall P: Recruitment of DNA replication and damage response proteins to viral replication centers during infection with NS2 mutants of Minute Virus of Mice (MVM). Virology. 2011 Feb 20; 2010 Dec 30. PMID: 21193212
- Cotmore SF, Hafenstein S, Tattersall P: Depletion of virion-associated divalent cations induces parvovirus minute virus of mice to eject its genome in a 3'-to-5' direction from an otherwise intact viral particle. J Virol. 2010 Feb; 2009 Dec 2. PMID: 19955311
- Kahn JS, Kesebir D, Cotmore SF, D'Abramo A Jr, Cosby C, Weibel C, Tattersall P: Seroepidemiology of human bocavirus defined using recombinant virus-like particles. J Infect Dis. 2008 Jul 1. PMID: 18491974
- Cotmore SF, Gottlieb RL, Tattersall P: Replication initiator protein NS1 of the parvovirus minute virus of mice binds to modular divergent sites distributed throughout duplex viral DNA. J Virol. 2007 Dec; 2007 Sep 26. PMID: 17898054
- Paglino J, Burnett E, Tattersall P: Exploring the contribution of distal P4 promoter elements to the oncoselectivity of Minute Virus of Mice. Virology. 2007 Apr 25; 2006 Dec 18. PMID: 17175002
- Cotmore SF, Tattersall P: Parvoviral host range and cell entry mechanisms. Adv Virus Res. 2007. PMID: 17765706
- Burnett E, Cotmore SF, Tattersall P: Segregation of a single outboard left-end origin is essential for the viability of parvovirus minute virus of mice. J Virol. 2006 Nov; 2006 Aug 23. PMID: 16928767
- Ruiz Z, D'Abramo A Jr, Tattersall P: Differential roles for the C-terminal hexapeptide domains of NS2 splice variants during MVM infection of murine cells. Virology. 2006 Jun 5; 2006 Feb 28. PMID: 16504232
- Farr GA, Cotmore SF, Tattersall P: VP2 cleavage and the leucine ring at the base of the fivefold cylinder control pH-dependent externalization of both the VP1 N terminus and the genome of minute virus of mice. J Virol. 2006 Jan. PMID: 16352540
- Farr GA, Zhang LG, Tattersall P: Parvoviral virions deploy a capsid-tethered lipolytic enzyme to breach the endosomal membrane during cell entry. Proc Natl Acad Sci U S A. 2005 Nov 22; 2005 Nov 11. PMID: 16284249
- D'Abramo AM Jr, Ali AA, Wang F, Cotmore SF, Tattersall P: Host range mutants of Minute Virus of Mice with a single VP2 amino acid change require additional silent mutations that regulate NS2 accumulation. Virology. 2005 Sep 15. PMID: 16039688
- Cotmore SF, Tattersall P: Encapsidation of minute virus of mice DNA: aspects of the translocation mechanism revealed by the structure of partially packaged genomes. Virology. 2005 May 25. PMID: 15866075
- Wollmann G, Tattersall P, van den Pol AN: Targeting human glioblastoma cells: comparison of nine viruses with oncolytic potential. J Virol. 2005 May. PMID: 15857987
- Cotmore SF, Tattersall P: Genome packaging sense is controlled by the efficiency of the nick site in the right-end replication origin of parvoviruses minute virus of mice and LuIII. J Virol. 2005 Feb. PMID: 15681430
- Farr GA, Tattersall P: A conserved leucine that constricts the pore through the capsid fivefold cylinder plays a central role in parvoviral infection. Virology. 2004 Jun 1. PMID: 15193920
- Palmer GA, Brogdon JL, Constant SL, Tattersall P: A nonproliferating parvovirus vaccine vector elicits sustained, protective humoral immunity following a single intravenous or intranasal inoculation. J Virol. 2004 Feb. PMID: 14722265
- Burnett E, Tattersall P: Reverse genetic system for the analysis of parvovirus telomeres reveals interactions between transcription factor binding sites in the hairpin stem. J Virol. 2003 Aug. PMID: 12885883
- Cotmore SF, Tattersall P: Resolution of parvovirus dimer junctions proceeds through a novel heterocruciform intermediate. J Virol. 2003 Jun. PMID: 12743281