William E. Goldman, Ph.D.

goldman@med.unc.edu 519A Mary Ellen Jones CB# 7290 Chapel Hill, NC 27599-7290 919.966.9580

Research

Successful respiratory pathogens must be able to respond swiftly to a wide array of sophisticated defense mechanisms in the mammalian lung. In histoplasmosis, macrophages — a first line of defense in the lower respiratory tract — are effectively parasitized by Histoplasma capsulatum. This process depends on virulence factors produced as this "dimorphic" fungus undergoes a temperature-triggered conversion from a saprophytic mold form to a parasitic yeast form. One such molecule is a calcium-binding protein (CBP) that is secreted preferentially by the yeast form and is essential for Histoplasma virulence. The experiments to unravel CBP structure and function have relied heavily on our development of a telomeric shuttle plasmid that has been used for complementation cloning, gene disruptions, RNA interference, and reporter gene constructs. In addition, random insertional mutagenesis and transcriptional profiling with microarrays are helping us identify and characterize genes involved in the regulation of CBP1 expression.

Another yeast phase-specific product of H. capsulatum is alpha-(1,3)-glucan, a cell wall polysaccharide that is associated with virulence in a variety of fungal pathogens. We have taken two approaches to study alpha-(1,3)-glucan: the first is a forward genetics strategy, using Agrobacterium-mediated insertional mutagenesis, to identify genes implicated in the regulation, synthesis, and processing of this polysaccharide. The second approach uses reverse genetics, combining fungal gene disruption with mammalian RNA-interference, to study the genes involved in production of and response to alpha-(1,3)-glucan. This work has revealed that alpha-(1,3)-glucan on the surface of Histoplasma yeasts masks recognition of the underlying beta-glucan by dectin-1, a macrophage pattern-recognition receptor that is critical in the innate immune response to fungi.

Yersinia pestis also displays two temperature-regulated lifestyles, depending on whether it is colonizing a flea or mammalian host. Inhalation by humans leads to a rapid and overwhelming disease, and we are trying to understand the development of pneumonic plague by studying genes that are activated during the stages of pulmonary colonization. We have generated an oligonucleotide-based microarray corresponding to all of the predicted genes of a pneumonic plague isolate, and this has been used for in vivo transcriptional profiling during experimental infection. We have also developed and characterized a mouse model for studying the pathological and immunological changes during the progression of pneumonic plague. This model system has revealed two sharply contrasting phases to the syndrome: the first phase of infection features rapid bacterial proliferation in the lung, but almost no inflammatory response, symptoms, or pathology; the second phase, starting at approximately 36 hours post-inoculation, is marked by inflammation and pneumonia that lead quickly to death. The utility of this model was highlighted in a recent study that demonstrated how a plasminogen-activating protease (encoded by the pla gene of Y. pestis) is essential for development of the second phase of this disease. We have now initiated a more comprehensive research plan, combining forward genetics with microarray technology, to determine what other bacterial genes are specifically required for the development of these two phases of pneumonic plague.

We are also continuing studies of one of the virulence factors of Bordetella pertussis: tracheal cytotoxin (TCT) is a released fragment of peptidoglycan that causes pulmonary inflammation in pertussis (whooping cough). TCT and endotoxin synergistically trigger respiratory epithelial production of nitric oxide, causing ciliated cell damage that corresponds to the well-known airway cytopathology of pertussis. A variety of host receptor systems have been shown to recognize peptidoglycan fragments, and some – such as the PGRP family – are evolutionarily conserved from flies to mammals. Depending on the relationship between bacteria and host, the results of exposure to TCT can be beneficial or pathological. Our current work is aimed at understanding host responses to TCT that include epithelial defense, cytopathology, and remodeling.

Publications

Kügler, S., T.S. Sebghati, L.G. Eissenberg, and W.E. Goldman. 2000. Phenotypic variation and intracellular parasitism by Histoplasma capsulatum. Proceedings of the National Academy of Sciences U.S.A. 97:8794-8798.

Sebghati, T.S., J.T. Engle, and W.E. Goldman. 2000. Intracellular parasitism by Histoplasma capsulatum: Fungal virulence and calcium dependence. Science 290:1368-1372.

Flak, T.A., L.N. Heiss, J.T. Engle, and W.E. Goldman. 2000. Synergistic epithelial responses to endotoxin and a naturally occurring muramyl peptide. Infection and Immunity 68:1235-1242.

Kügler, S., B. Young, V.L. Miller, and W.E. Goldman. 2000. Monitoring phase-specific gene expression in Histoplasma capsulatum with telomeric GFP fusion plasmids. Cellular Microbiology 2:537-547.

Magrini, V., and W.E. Goldman. 2001. Molecular mycology: A genetic toolbox for Histoplasma capsulatum. Trends in Microbiology 9:541-546.

Liu, H., T.R. Cottrell, L.M. Pierini, W.E. Goldman, and T.L. Doering. 2002. RNA interference in the pathogenic fungus Cryptococcus neoformans. Genetics 60:463-470.

Kaneko, T., W.E. Goldman, P. Mellroth, H. Steiner, K. Fukase, S. Kusumoto, W. Harley, A. Fox, D. Golenbock, and N. Silverman. 2004. Monomeric and polymeric gram-negative peptidoglycan but not purified LPS stimulate the Drosophila IMD pathway. Immunity 20:637-649.

Magrini, V., W.C. Warren, J. Wallis, W.E. Goldman, J. Xu, E.R. Mardis, and J.D. McPherson. 2004. Fosmid-based physical mapping of the Histoplasma capsulatum genome. Genome Research 14:1603-1609.

Rappleye, C.A., J.T. Engle, and W.E. Goldman. 2004. RNA interference in Histoplasma capsulatum demonstrates a role for alpha-(1,3)-glucan in virulence. Molecular Microbiology 53:153-165.

Koropatnick, T.A., J.T. Engle, M.A. Apicella, E.V. Stabb, W.E. Goldman, and M.J. McFall-Ngai. 2004. Microbe factor-mediated development in a host-bacterial mutualism. Science 306:1186-1188.

Mellroth, P., J. Karlsson, J. Håkansson, N. Schultz, W.E. Goldman, and H. Steiner. 2005. Ligand induced dimerization of Drosophila peptidoglycan recognition proteins. Proceedings of the National Academy of Sciences U.S.A. 102:6455-6460.

Lathem, W.W., S.D. Crosby, V.L. Miller, and W.E. Goldman. 2005. Progression of primary pneumonic plague: A mouse model of infection, pathology, and bacterial transcriptional activity. Proceedings of the National Academy of Sciences U.S.A. 102:17786-17791.

Swaminathan, C.P., P.H. Brown, A. Roychowdhury, Q. Wang, R. Guan, N. Silverman, W.E. Goldman, G.-J. Boons, and R.A. Mariuzza. 2006. Dual strategies for peptidoglycan discrimination by peptidoglycan recognition proteins (PGRPs). Proceedings of the National Academy of Sciences U.S.A. 103:684-689.

Lim J.H., M.S. Kim, H.E. Kim, T. Yano, Y. Oshima, K. Aggarwal, W.E. Goldman, N. Silverman, S. Kurata, and B.H. Oh. 2006. Structural basis for preferential recognition of diaminopimelic acid-type peptidoglycan by a subset of peptidoglycan recognition proteins. Journal of Biological Chemistry 281:8286-8295.

Rappleye, C.A., and W.E. Goldman. 2006. Defining virulence genes in the dimorphic fungi. Annual Review of Microbiology 60:281-303.

Mielcarek, N., A.-S. Debrie, D. Raze, J. Bertout, A. Ben Younes, J. Engle, W.E. Goldman, and C. Locht. 2006. Live attenuated Bordetella pertussis as a highly efficient single-dose mucosal vaccine against whooping cough. PLoS Pathogens 2:e65.

Kaneko, T., T. Yano, K. Aggarwal, J.H. Lim, K. Ueda, Y. Oshima, C. Peach, D. Erturk-Hasdemir, W.E. Goldman, B.H. Oh, S. Kurata and N. Silverman. 2006. PGRP-LC and PGRP-LE play essential yet distinct roles in the Drosophila immune response to monomeric DAP-type peptidoglycan. Nature Immunology 7:715-723.

Cathelyn, J., S.D. Crosby, W.W. Lathem, W.E. Goldman, and V.L. Miller. RovA, a global regulator of Yersinia pestis, specifically required for bubonic plague. 2006. Proceedings of the National Academy of Sciences U.S.A. 103:13514-13519.

Marion, C.L., C.A. Rappleye, J.T. Engle, and W.E. Goldman. 2006. An alpha-(1,4)-amylase is essential for alpha-(1,3)-glucan production and virulence in Histoplasma capsulatum. Molecular Microbiology 62:970-983.

Cloud-Hansen, K.A., S.B. Peterson, E.V. Stabb, W.E. Goldman, M.J. McFall-Ngai, and J. Handelsman. 2006. Breaching the great wall: Peptidoglycan and microbial interactions. Nature Reviews Microbiology 4:710-716.

Lathem, W.W., P.A. Price, V.L. Miller, and W.E. Goldman. 2007. A plasminogen-activating protease specifically controls the development of primary pneumonic plague. Science 26:509-513.

Rappleye, C.A., L.G. Eissenberg, and W.E. Goldman. 2007. Histoplasma capsulatum alpha-(1,3)-glucan blocks recognition by the macrophage beta-glucan receptor. Proceedings of the National Academy of Sciences U.S.A. 104:1366-1370.

Rappleye, C.A., and W.E. Goldman. 2008. Fungal stealth technology. Trends in Immunology 29:18-24.

Beck, M.R., G.T. DeKoster, D.M. Hambly, M.L. Gross, D.P. Cistola, and W.E. Goldman. Structural features responsible for biological stability of Histoplasma’s virulence factor CBP. 2008. Biochemistry 47:4427-4438.

Yano, T., S. Mita, H. Ohmori, Y. Oshima, Y. Fujimoto, R. Ueda, H. Takada, W.E. Goldman, K. Fukase, N. Silverman, T. Yoshimori, S. Kurata. 2008. Autophagic control of listeria through intracellular innate immune recognition and autophagy in drosophila. Nature Immunology 9:908-916.

Adin, D.M, J.T. Engle, W.E. Goldman, M.J. McFall-Ngai, and E.V. Stabb. 2009. Mutations in ampG and lytic transglycosylase genes affect the net release of peptidoglycan monomers from Vibrio fischeri. Journal of Bacteriology 191:2012-2022.

Beck, M.R., G.T. DeKoster, D.P. Cistola, and W.E. Goldman. 2009. NMR structure of a fungal virulence factor reveals structural homology with mammalian saposin B. Molecular Microbiology 72:344-353.

Troll, J.V., D.M. Adin, A.M. Wier, N. Paquette, N. Silverman, W.E. Goldman, F.J. Stadermann, E.V. Stabb, and M.J. McFall-Ngai. 2009. Peptidoglycan induces loss of a nuclear peptidoglycan recognition protein during host tissue development in a beneficial animal-bacterial symbiosis. Cellular Microbiology 11:1114-1127.

Lehotzky, R.E., C.L. Partch, S. Mukherjee, H.L. Cash, W.E. Goldman, K.H. Gardner, and L.V. Hooper. 2010. Molecular basis for peptidoglycan recognition by a bactericidal C-type lectin. Proceedings of the National Academy of Sciences U.S.A. 107:7722-7727.

Weening, E.H., J.S. Cathelyn, G. Kaufman, M.B. Lawrenz, P. Price, W.E. Goldman, and V.L. Miller. 2011. The dependence of the Yersinia pestis capsule on pathogenesis is influenced by the mouse background. Infection and Immunity 79:644-652.

Bobrov, A.G., O. Kirillina, D.A. Ryjenkov, C.M. Waters, P.A. Price, J.D. Fetherson, D. Mack, W.E. Goldman, M. Gomelsky, and R.D. Perry. 2011. Systematic analysis of cyclic di-GMP signalling enzymes and their role in biofilm formation and virulence in Yersinia pestis. Molecular Microbiology 79:533-551.

Altura, M.A., E. Stabb, W. Goldman, M. Apicella, M.J. McFall-Ngai. 2011. Attenuation of host NO production by MAMPs potentiates development of the host in the squid-vibrio symbiosis. Cellular Microbiology 13:527-537.

Price, P.A., J. Jin, and W.E. Goldman. 2012. Pulmonary infection by Yersinia pestis rapidly establishes a permissive environment for microbial proliferation. Proceedings of the National Academy of Sciences U.S.A. 109:3083-3088.

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Accomplishments

Honors and Leadership Activities
1999              ASM Division D (Bacteria of Medical Importance) Lecturer
2000              Chair of FASEB Research Conference on Microbial Pathogenesis
1996-2001    Burroughs Wellcome Fund Scholar Award in Molecular Pathogenic Mycology
2002              Chair of Gordon Research Conference on Microbial Toxins and Pathogenicity
1998-2007    Director, Washington University Graduate Program in Molecular Microbiology
2002-present Fellow of the American Academy of Microbiology (AAM)

Other Professional Activities
1990-1994    Member, NIH Biological Sciences Study Section, Subcommittee 3
1998              Ad hoc member, NIAID Board of Scientific Counselors
1987-2001    Editorial Board, Infection & Immunity
1991-2002    Ad hoc reviewer, NIH Bacteriology and Mycology Study Section, Subcommittees 1 & 2
2003-2004    AAM Colloquia on new directions/issues in microbiology and genomics
2005              ASM-NIH Workshop on Basic Bacterial Research
2001-2008    Editor, Molecular Microbiology
2001, 2009    ASM General Meeting Colloquium Advisory Committee
2007-2009    Program Committee, ASM Biodefense and Emerging Diseases Research Meeting
1984-present 80 lectures at international conferences; 125 invited seminars at research institutes
1997-present Editorial Boards: Cell. Microbiol., Curr. Opin. Microbiol., Trends Microbiol., Eukaryotic Cell
2001-present "Faculty of 1000" Section Co-Head (Cellular Microbiology and Pathogenesis)
2004-present  ASM Conference Committee (Chair in 2012)
2005-2012     Advisory Committee, Burroughs Wellcome Fund
2011-present "Pearls" Editor, PLoS Pathogens