Life Science III 1007
Research Specialties: Bacterial pathogenesis focusing on Chlamydia. Chlamydia spp. are Gram negative, obligate intracellular bacterial pathogens which cause disease in humans as well as economically important domestic animals. These bacteria mainly infect the mucosal epithelial layer lining the lung, ocular, digestive, and genital tracts eliciting damage primarily in the form of immunopathology. In humans, C. trachomatis is responsible for trachoma (the leading cause of preventable infectious blindness worldwide) and is the leading cause of reported bacterial sexually transmitted infections (STIs) in the United States (and worldwide) with over 1.5 million cases reported in 2015 (CDC). Infections in women are of particular concern as they can lead to pelvic inflammatory disease, life-threatening ectopic pregnancy, or infertility. C. pneumoniae infects the respiratory tract causing pneumonia. While the incidence of chlamydial pneumonia is unknown, there are over 2 million reported cases of pneumonia in the United States each year (CDC). Furthermore, C. pneumoniae infections are a suspected risk factor for the development of cardiovascular disease and other chronic conditions. While antibiotics are relatively effective in treating chlamydial infections, treatment failure is a growing concern and there is a need for the discovery of new antibiotic targets. Concurrently, the high percentage of asymptomatic chlamydial infections (≥50% of STIs are asymptomatic), and the link between these infections and chronic disease, illustrates the urgent need to develop vaccines and anti-infectives.
PhD, 2006, University of Pittsburgh School of Medicine
My research is broadly focused on how bacterial physiology, using Chlamydia as a model pathogen, impacts virulence based on the premise that most bacterial pathogens must be able to obtain nutrients, grow, and divide to cause disease. These bacteria transition between two distinct developmental forms, the environmentally stable infectious form known as the elementary body (EB) and the intracellular replicative form known as the reticulate body (RB). For successful reproduction, the EB must bind and become internalized in a eukaryotic cell, modify host-vesicular trafficking pathways, differentiate into an RB, transition back into an EB, and finally exit from the host cell into the environment. Despite this complicated developmental cycle and its essentiality to infection, the mechanisms regulating and carrying out morphogenesis remain largely undefined. Recently, global bacterial Ser/Thr/Tyr protein phosphorylation has become an increasingly appreciated method for regulating protein function in bacteria. Chlamydia spp. encode two validated Ser/Thr kinases and our group recently validated a PP2C protein phosphatase. However, the extent of phosphorylation in Chlamydia throughout development and the role of phosphorylation in chlamydial physiology and virulence is ill-defined.
As a first step in determining the role of phosphorylation in chlamydial biology, we mapped the phosphoproteome of Chlamydia caviae GPIC EB and RB developmental forms. This research had three key findings: 1) that C. caviae are capable of global Ser/Thr/Tyr protein phosphorylation (phosphorylating >4% of its proteome), 2) proteins phosphorylated in C. caviae are highly conserved throughout Chlamydia spp. suggestive of a conserved chlamydial phosphoproteome, and 3) that protein phosphorylation appears to be more prevalent in EBs (3-fold more phosphoproteins) than in RBs. Phosphoprotein profiles of other Chlamydia spp. demonstrate similar phosphoprotein numbers and developmental skewing compared to C. caviae. Collectively, these data led us to hypothesize that phosphorylation plays a key role in regulating morphogenesis. The extent, conservation, and developmental skew of protein phosphorylation provide support for further studies to discern the importance and consequences of protein phosphorylation in Chlamydia. In addition, work by others and our research team has shown that chemical inhibition of kinase and phosphatase activity reduces bacterial growth indicating that these proteins might serve as novel antimicrobial targets.
Consequently, my research focuses on 1) elucidating the function and regulation of the chlamydial protein kinases and phosphatases during bacterial growth, 2) characterizing the phosphorylation-controlled chlamydial partner switching mechanism, and 3) assessing the role of phosphorylation in regulating key physiological processes, including metabolism, found to be differentially phosphorylated in EBs versus RBs. In addition, my research seeks to expand upon the more recent breakthrough in chlamydial genetics through the development of tools to increase the methods available to genetically manipulate Chlamydia. These projects will provide important insights into the biology of Chlamydia, identify pathways that can be targeted to disrupt its pathogenic abilities, and will generate tools useful for the design of live-attenuated strains for vaccine studies.
Articles in Professional Journals
- Pais, S.V., Key, C.E., Borges, V., Pereira, I.S., Gomes, J.P., Fisher, D.J., and Mota, L.J. (2019) CteG is a Chlamydia trachomatis effector protein that associates with the Golgi complex of infected host cells. Scientific Reports. PMID: 30992493.
- Shaw, J.H., Key, C.E., Snider, T.A., Shaw, E., Fisher, D.J., and Lutter, E.I. (2018) Genetic inactivation of Chlamydia trachomatis inclusion membrane protein CT228 alters MYPT1 recruitment, extrusion production and longevity of infection. Frontiers in Cellular and Infection Microbiology. DOI: 10.3389/fcimb.2018.00415. Link
- Wood, N.A., Chung, K.Y., Blocker, A.M., Rodrigues de Almeida, N., Conda-Sheridan, M., Fisher, D.J., and Ouellette, S.P. (2018) Initial Characterization of the Two ClpP Paralogs of Chlamydia trachomatis Suggests Unique Functionality for Each. Journal of Bacteriology. PMID: 30396899. Link
- Claywell, J.E., Matschke, L.M., Plunkett, K.N., and Fisher, D.J. (2018) Inhibition of the Protein Phosphatase CppA Alters Development of Chlamydia trachomatis. Journal of Bacteriology. DOI: 10.1128/JB.00419-18. PMID: 30038048 Link
- Cossé, M.M., Barta, M.L., Fisher, D.J., Oesterlin, L.K., Niragire, B., Perrinet, S., Millot, G.A., Hefty, P.S., and Subtil, A. (2018) The Loss of Expression of a Single Type 3 Effector (CT622) Strongly Reducs Chlamydia trachomatis Infectivity and Growth. Frontiers in Cellular and Infection Microbiology. 8:145. PMID: PubMed Link
- Gautama, D., Umagiliyagea, A.L., Dhitala, R., Joshia, P., Watson, D.G., Fisher, D.J., and Choudhary, R. (2017) Nonthermal pasteurization of tender coconut water using a continuous flow coiled UV reactor. LWT - Food Science and Technology. 83: 127-137. DOI: 10.1016/j.lwt.2017.05 .008 Link
- Illingworth, M., Hooppaw, A.J., Ruan, L., Fisher, D.J. and Chen, L. (2017) Biochemical and genetic analysis of the Chlamydia trachomatis GroEL chaperonins. Journal of Bacteriology. PMID: 28396349. PubMed Link
- Umagiliyage, A.L., Becerra-Mora, N., Kohli, P., Fisher, D.J., and Choudhary, R. (2017) Antimicrobial efficacy of liposomes containing d-limonene and its effect on the storage life of blueberries. Postharvest Biology and Technology. Link
- Claywell, J.E., Matschke, L.M., and Fisher, D.J. (2016) The impact of protein phosphorylation on chlamydial physiology. Frontiers in Cellular and Infection Microbiology. 6:197. Link
- Key, C.E. and Fisher, D.J. (2016) Use of Group II Intron Technology for Targeted Mutagenesis in Chlamydia trachomatis. Methods in Molecular Biology. 1498:163-177. PMID: 27709575 PubMed Link
- Claywell, J.E. and Fisher, D.J. (2016) CTL0511 from Chlamydia trachomatis is a Type 2C Protein Phosphatase (PP2C) with Broad-Substrate Specificity. Journal of Bacteriology. PMID: 27114464 PubMed Link
- Zhang, H., Wang, Q., Fisher, D.J., Cai, M., Chakravartty, V., Ye, H., Li, P., Solbiati, J.O., Feng, Y. (2016) Deciphering a unique biotin scavenging pathway with redundant genes in the probiotic bacterium Lactococcus lactis. Scientific Reports. 6:25680. PMID: 27161258 PubMed Link
- Hooppaw, A.J. and Fisher, D.J.. (2015) A Coming of Age Story: Chlamydia in the Post-Genetic Era. Infection and Immunity. Accepted. (Invited Review). PMID: 26667838 PubMed Link
- Lowden, N.M., Yeruva, L., Johnson, C.M., Bowlin, A.K., Fisher, D.J. 2015. Use of aminoglycoside 3′ adenyltransferase as a selection marker for Chlamydia trachomatis intron-mutagenesis and in vivo intron stability. BMC Research Notes. 8:570. Link
- Thompson, C.C., Griffiths, C., Nicod, S.S., Lowden, N.M., Wigneshweraraj, S., Fisher, D.J., McClure, M.O. 2015. The Rsb phosphoregulatory network controls availability of the primary sigma factor in Chlamydia trachomatis and influences the kinetics of growth and development. PLoS Pathogens. 11(8): e1005125. Link
- Fisher, D.J., Adams, N.E., and Maurelli, A.T. 2015. Phosphoproteomic analysis of the Chlamydia caviae elementary body and reticulate body forms. Microbiology 161: 1648-1658. PMID: 25998263. PubMed link
- Ma, M., Gurjar, A., Theoret, J.R., Garcia, J.P., Beingesser, J., Freeman, J.C., Fisher, D.J., McClane, B.A., and Uzal, F.A. 2014. Synergistic effects of Clostridium perfringens enterotoxin and beta toxin in rabbit small intestinal loops. Infect Immun. 82(7): 2958-2970. PMID: 24778117. PubMed link
- Johnson, C.M and Fisher, D.J. 2013. Site-specific, insertional inactivation of incA in Chlamydia trachomatis using a Group II intron. PLoS ONE 8(12): e83989. PMID: 24391860. Link
- Fisher, D.J., Fernández, R.E., and Maurelli, A.T. 2013. Chlamydia trachomatis transports NAD via the Npt1Ct ATP/ADP translocase. Journal of Bacteriology. 195(15): 3381-6. PMID: 23708130. PubMed link
- Garcia, J.P, Beingesser, J., Fisher, D.J., Sayeed, S., McClane, B.A., Posthaus, H., and Uzal, F.A. (2012) The effect of Clostridium perfringens type C strain CN3685 and its isogenic beta toxin null mutant in goats. Vet Microbiol. 157(3-4): 412-9. PMID: 22296994. PubMed Link
- *Bliven, K.A., *Fisher, D.J., Maurelli, A.T. 2012. Characterization of the activity and expression of arginine decarboxylase in human and animal Chlamydia pathogens. FEMS Microbiology Letters 337:140-6. PMID:23043454. PubMed link *These authors contributed equally to this work.
- Fisher, D.J., Fernández, R.E., Adams, N.E., and Maurelli, A.T. 2012. Uptake of Biotin by Chlamydia Spp. Through the Use of a Bacterial Transporter (BioY) and a Host-Cell Transporter (SMVT). PLOS ONE. Link
- Binet, R., Fernández, R.E., Fisher, D.J. and Maurelli, A.T. 2011. Identification and Characterization of theChlamydia trachomatis L2 S-Adenosyl Methionine / S-Adenosyl Homocysteine transporter. mBio. May 10;2(3):e00051-11. doi: 10.1128/mBio.00051-11. PubMed link
- Giles, T.N., Fisher, D.J. and Graham, D.E. 2009. Independent inactivation of arginine decarboxylase genes by nonsense and missense mutations led to pseudogene formation in Chlamydia trachomatis serovar L2 and D strains. BMC Evolutionary Biology. 9:166. PMCID: PMC2720952 PubMed link
- Uzal, F.A., Saputo, J., Sayeed S, Vidal, J.E., Fisher, D.J., Poon, R., Adams, V., Fernandez-Miyakawa, M.E., Rood, J.I. and McClane, B.A. 2009. Development and application of new mouse models to study the pathogenesis of Clostridium perfringens type C Enterotoxemias. Infection and Immunity. 77(12): 5291-9. PMCID: PMC2786445 PubMed link
- Sayeed, S., Uzal, F.A., Fisher, D.J., Saputo, J., Vidal, J.E., Chen, Y., Gupta, P., Rood, J.I. and McClane, B.A. 2008. Beta toxin is essential for the intestinal virulence of Clostridium perfringens type C disease isolate CN3685 in a rabbit ileal loop model. Molecular Microbiology. 67(1): 15-30. (Figure from paper used for issue cover) PubMed link
- Uzal, F.A., Fisher, D.J., Saputo, J., Sayeed, S., McClane, B.A., Songer, G., Trinh, H.T., Fernandez-Miyakawa, M.E. and Gard, S. 2008. Ulcerative enterocolitis in two goats associated with enterotoxin- and beta2 toxin-positive Clostridium perfringens type D. Journal of Veterinary Diagnostic Investigation. 20(5):668-72. Link
- Shivaprasad, H.L. Uzal, F., Kokka, R., Fisher, D.J., McClane, B.A. and Songer, A.G. 2008. Ulcerative enteritis-like disease associated with Clostridium perfringens type A in bobwhite quail (Colinus virginianus). Avian Diseases. 52(4):635-40. PubMed link
- Fernandez-Miyakawa, M.E., Sayeed, S., Fisher, D.J., Poon, R. Adams, V., Rood, J.I., McClane, B.A., Saputo, J. and Uzal, F.A. 2007. Development and application of a mouse oral challenge model for studying Clostridium perfringens type D infection. Infection and Immunity. 75(9): 4282-8. PMCID: PMC1951146 PubMed link
- Chen, Y., Caruso, L., McClane, B., Fisher, D. and Gupta, P. 2007. Disruption of a toxin gene by introduction of a foreign gene into the chromosome of Clostridium perfringens using targetron-induced mutagenesis. Plasmid. 58(2): 182-9. PMCID: PMC2034400 PubMed link
- Miyakawa, M.E., Saputo, J., Leger, J.S., Puschner, B., Fisher, D.J., McClane, B.A. and Uzal, F.A. 2007. Necrotizing enterocolitis and death in a goat kid associated with enterotoxin (CPE)-producing Clostridium perfringens type A. Canadian Veterinary Journal. 48(12): 1266-9. PMCID: PMC2081994 PubMed link
- Fernandez-Miyakawa, M.E., Fisher, D.J., Poon, R., Sayeed, S., Adams, V., Rood, J.I., McClane, B.A. and Uzal, F.A. 2007. Both epsilon-toxin and beta-toxin are important for the lethal properties of Clostridium perfringenstype B isolates in the mouse intravenous injection model. Infection and Immunity. 75(3): 1443-1452. PMCID: PMC1828578 PubMed link
- Crespo, R., Fisher, D.J., Shivaprasad, H.L., Fernandez-Miyakawa, M.E. and Uzal, F.A. 2007. Toxinotypes ofClostridium perfringens isolated from sick and healthy avian species. Journal of Veterinary Diagnostic Investigation. 19(3): 329:33. PubMed link
- Fisher, D.J., Fernandez-Miyakawa, M.E., Sayeed, S., Poon, R., Adams, V., Rood, J.I., Uzal, F.A. and McClane, B.A. 2006. Dissecting the lethality contributions of Clostridium perfringens type C toxins to lethality in the mouse intravenous injection model. Infection and Immunity, 74(9): 5200-5210. PMCID: PMC1594841 PubMed link
- Myers, G.S.A., Rasko, D.A., Cheung, J.K., Ravel, J., Seshadri, R.,DeBoy, R.T., Ren, Q., Varga, J., Awad, M.M., Brinkac, L.M., Daugherty, S.C., Haft, D.H., Dodson, R.J., Madupu, R., Nelson, W.C., Rosovitz, M.J., Sullivan, S.A., Khouri, H., Dimitrov, G.I., Watkins, K.L., Mulligan, S., Benton, J., Fisher, D.J., Atkins, H.S., Hiscox, T., Jost, H., Billington, S.J., Songer, J.G., McClane, B.A., Titball, R.W., Rood, J.I., Melville, S. and Paulsen, I.T. 2006. Skewed Genomic Variability in Strains of the Toxigenic Bacterial Pathogen, Clostridium perfringens. Genome Research, 16: 1031-1040. PMCID: PMC1524862 PubMed link
- Miyamoto, K., Fisher, D.J., Li, J., Sayeed, S., Akimoto, S. and McClane, B.A. 2006. Complete sequencing and diversity analysis of the enterotoxin-encoding plasmids in Clostridium perfringens type A nonfoodborne human gastrointestinal disease isolates. Journal of Bacteriology, 188(4):1585-1598. PMCID: PMC1367241 PubMed link
- Chen, Y., McClane, B., Fisher, D.J., Rood, J. and Gupta, P. 2005. Construction of an Alpha Toxin Gene Knockout Mutant of Clostridium perfringens Type A using a Mobile Group II Intron. Applied and Environmental Microbiology, 71(11):7542-7. PMCID: PMC1287605 PubMed link
- Sayeed, S., Fernandez-Miyakawa, M.E., Fisher, D.J., Adams, V., Poon, R., Rood, J.I., Uzal, F.A. and McClane, B.A. 2005. Epsilon toxin is required for most Clostridium perfringens type D vegetative culture supernatants to cause lethality in the mouse intravenous injection model. Infection and Immunity, 73(11):7413-21. PMCID: PMC1273886 PubMed link
- Fisher, D.J., Miyamoto, K., Sarker, M.R., Harrison, B. And McClane, B.A. 2005. Association of beta2 toxin production with Clostridium perfringens type A human gastrointestinal disease isolates carrying a plasmid enterotoxin gene. Molecular Microbiology 56(3):747-762. PubMed link
- Smedley III, J.G., Fisher, D.J., Sayeed, S., Chakrabarti, G. and McClane, B.A. 2004. The enterotoxins ofClostridium perfringens. Reviews in Physiology, Biochemistry and Pharmacology 72(12):6914-23. (Invited Review) PubMed link
- Zhou, Z., Fisher, D., Spidel, J., Greenfield, J., Patson, B., Fazal, A., Wigal, C., Moe, O.A. and Madura, J.D. 2003. Kinetic and docking studies of the interaction of quinones with the quinone reductase active site. Biochemistry 42(7):1985-94. PubMed link