Microbiology and Immunology
Education and Training
- Widener University, BS, Biology, 1998
- University of Delaware, MS, Entomology/Applied Ecology, 2001
- Texas A&M University, PhD, Entomology, 2005
- Virginia Bioinformatics Institute (Virginia Tech), Post-Doctoral Fellow, 2006-2008
- Virginia Bioinformatics Institute (Virginia Tech), Senior Research Associate, 2009-2013
Dr. Gillespie is an evolutionary biologist with broad interests in organismal and molecular evolution. The major focus of his current research is deciphering the mechanisms by which obligate intracellular species of Rickettsiales (Alphaproteobacteria) invade, survive and replicate within eukaryotic cells.
In research funded by the National Institutes of Health, Dr. Gillespie utilizes phylogenetics, comparative genomics and bioinformatics to guide experimental studies on various pathogenic species of Rickettsia and their associated arthropod vectors. His early research resulted in the reclassification of Rickettsia species and the identification of many lineage-specific pathogenicity factors. Through years of intense scrutinization of dozens of diverse rickettsial genomes, Dr. Gillespie and colleagues have described a large, dynamic mobilome for Rickettsia species, resulting in the identification of integrative conjugative elements as the vehicles for seeding Rickettsia genomes with many of the factors underlying obligate intracellular biology and pathogenesis. Via an iterative process of genome sequencing, phylogenomics, bioinformatics, and classical molecular biology and microbiology, Dr. Gillespie continues to lead and assist research projects on the characterization of rickettsial gene and protein function, as well as the description of cell envelope glycoconjugates, such as peptidoglycan and lipopolysaccharide.
A particular focal area of Dr. Gillespie’s research is the Rickettsia secretome, which includes the secretion systems and their cognate substrates, many of which directly engage arthropod and vertebrate molecules throughout the rickettsial infection process. Dr. Gillespie led a study that identified across all genera of Rickettsiales the composition of an enigmatic type IV secretion system (T4SS), termed rvh (Rickettsiales vir homolog). Several collaborative efforts have begun elucidating the odd architecture of the rvh T4SS, as well as the protein substrates (e.g., RalF, RARP-2, and Risk1) it translocates into host cells during infection. A recent analysis of rickettsial metagenomes provided substantial insight on the evolution of effector architectures.
Evolutionary Biology, Rickettsiology, Rickettsia, Microbiology, Molecular Biology, Genetics, Bioinformatics, Phylogenetics, Phylogenomics, Metabolomics, Structural Biology, Lateral Gene Transfer, Type IV Secretion Systems, Secretome, Pathogenesis, Genome Sequencing, Arthropods, Ticks, Fleas, Body Lice, Epidemic Typhus, Murine Typhus, Rocky Mountain Spotted Fever, Transitional Group rickettsiae, Wolbachia
Yang, H., Verhoeve, V.I., Chandler, C.E., Nallar, S., Snyder, G.A., Salje, J., Ernst, R.K., Gillespie, J.J. (2023) Structural determination of Rickettsia lipid A without chemical extraction confirms shorter acyl chains in later-evolving Spotted Fever Group pathogens. View.
Giengkam, S., Kullapanich, C. Wongsantichon, J., Adcox H.E., Gillespie, J.J., Salje, J. (2023) Orientia tsutsugamushi: analysis of the mobilome of a highly fragmented and repetitive genome reveals ongoing lateral gene transfer in an obligate intracellular bacterium. View.
Verhoeve, V.I., Lehman, S.S., Driscoll, T.P., Beckmann, J.F., Gillespie, J.J. (2023)
Beckmann, J.F., Gillespie, J.J., Tauritz, D. (2023) Modelling emergence of Wolbachia toxin-antidote protein functions with an evolutionary algorithm
Gillespie, J.J., Salje, J. (2023) Orientia and Rickettsia: different flowers from the same garden. Current Opinion in Microbiology 74: 102318.
Verhoeve, V.I., Gillespie, J.J. (2022)
Verhoeve VI, Brammer JA, Driscoll TP, Kambouris AR, Rasko DA, Cross AS, Gillespie, J.J. (2022) Genome sequencing of Pseudomonas aeruginosa strain M2 illuminates traits of an opportunistic pathogen of burn wounds. G3 12.
Verhoeve V.I., Fauntleroy T.D., Risteen R.G., Driscoll, T.P., Gillespie, J.J. (2022)
Atwal, S., Chuenklin, S., Bonder, E.M., Flores, J., Discovery of a Diverse Set of Bacteria That Build Their Cell Walls without the Canonical Peptidoglycan Polymerase aPBP. mBio. e0134221.
Guillotte, M.L., Chandler, C.E., Verhoeve, V.I., mSphere 6: e00184-21.
Gillespie, J.J., Joardar, V., Williams, K.P., Driscoll, T., Hostetler, J.B., Nordberg, E.K., Shukla, M., Walenz, B., Hill, C.A., Nene, V.M., Azad, A.F., Sobral, B.W. & Caler, E. (2012) A Rickettsia genome overrun by mobile genetic elements provides insight into the acquisition of genes characteristic of an obligate intracellular lifestyle. Journal of Bacteriology 194: 376-394.
Gillespie, J.J., Beier, M.S., Rahman, M.S., Ammerman, N.C., Purkayastha, A., Shallom, J.M., Sobral, B.S., Azad, A.F. (2007) Plasmids and rickettsial evolution: insight from Rickettsia felis. PLoS ONE 2: e266.
Additional Publication Citations
Dr. Gillespie's publications and related information can be found on the following web sites:
Sequencing of the deer tick (Ixodes scapularis) genome revealed the nature of a Rickettsia endosymbiont that is frequently found in deer tick populations. Genomic analysis of this species, now called Rickettsia buchneri, unearthed a large and diverse cache of mobile genetic elements. Prominent among these is a distinct integrative and conjugative element named RAGE (Rickettsiales Amplified Genetic Element). Dr. Gillespie and colleagues determined that RAGEs are the vehicles for seeding Rickettsia genomes with many of the factors underlying obligate intracellular biology and pathogenesis. Prior analyses discovered substantial diversity among R. buchneri RAGEs, both within individual ticks as well as in the I. scapularis population at large, indicating that RAGEs are actively mobilizing across Rickettsia species. Dr. Gillespie and a team led by colleague Dr. Jeanne Salje (University of Cambridge) recently determined that RAGEs are also actively transferring between genomes of the agent of Scrub Typhus, Orientia tsutsugamushi. Rickettsia and Orientia species are closely related rickettsial lineages that differ markedly in their biology and pathogenesis, allowing for many key insights through comparative analyses.
This RAGE-mediated lateral gene transfer has steered research and discovery in three main focal areas:
1. Virulence Factors
The search for Rickettsia secreted effectors that target host cell molecules begins with detecting probable products of lateral gene transfer (e.g., piggybacking on RAGEs, phylogenetic signal discordant with “housekeeping” genes) that may contain eukaryotic-like characteristics (e.g., ankyrin repeats, Sec7-like domains, phosphatidylinositol kinase motifs). Dr. Gillespie and colleagues then aim to identify the rickettsial secretion pathway involved in translocating such effectors. A major focus is the rvh T4SS, wherein effectors must bind the rvh coupling protein (RvhD4) prior to secretion. Several approaches are used to capture RvhD4-effector interactions, with subsequent characterization of effectors using in vitro and in vivo experiments. The effectors RalF, RARP-2 and Risk1 have been shown to be translocated into host cells by Rickettsia typh during infection, and other candidate effectors are currently being characterized.
Many of the RAGE cargo genes encode for proteins that are seemingly important for intracellular survival (e.g., osmoregulatory transporters, lipopolysaccharide-modifying enzymes, peptidoglycan recycling factors) and host-dependent metabolism (stringent response components, metabolite transporters, metabolic enzymes). As rickettsiae are subject to reductive genome evolution, the RAGEs appear to offset this deleterious process by shuttling across the rickettsial mobilome genes that are critical for intracellular parasitism. The importance for maintaining genes underpinning metabolism was realized through reconstructing the Rickettsia-host metabolic network, which revealed 51 host cell metabolites required to compensate the patchwork rickettsial metabolism. Dr. Gillespie and colleagues are studying the novel mechanisms utilized by rickettsiae to pilfer host metabolites from the eukaryotic cytoplasm. A major focus entails solving how rickettsiae steal amino sugars to supply building blocks for cell envelope synthesis (rickettsiae lack glycolysis and have quite a sweet tooth!).
3. Reproductive Parasitism
RAGEs have also been shown to shuttle genes implicated in reproductive parasitism (RP), which is the ability of microbial parasites to influence the sexual reproduction of their hosts (via processes such as male-killing, feminization, parthenogenesis, and cytoplasmic incompatibility, or CI). Parasites induce RP to drive their retention in host populations via the female germline. RP, particularly CI, is well-studied for Wolbachia parasites, which are close cousins to Rickettsia species and infect over half the world’s described arthropod species, as well as many nematode species. While the molecular basis for Wolbachia-mediated CI was discovered by Wolbachia researchers (classical toxin-antidote operons), Dr. Gillespie and colleagues identified divergent orthologs to these factors in a range of different intracellular parasites, including human pathogens (i.e., Rickettsia felis, Rickettsia gravesii and Orientia tsutsugamushi). Current studies are focused on elucidating the mechanisms of RP induction by these diverse toxin-antidote operons, as well as identifying novel reproductive parasites of the cat flea, a vector that transmits pathogenic rickettsiae that scourge the homeless population. Dr. Gillespie collaborates with Dr. John Beckmann, one of the pioneers for discovering the genetic basis for CI. A recent study developed a model to better understand the evolution of CI genes in arthropod populations. Furthermore, the team is currently characterizing CI-like genes of Orientia tsutsugamushi, which induces parthenogenesis in trombiculid mites.
Genome sequencing of the cat flea (Ctenocephalides felis) revealed a bizarre “genome in flux” characterized by inordinate copy number variation (~38% of proteins) and a broad range of genome size estimates (433-551 Mb) for individual fleas. Surprisingly, the flea genome exhibits neither inflation due to rampant gene duplication nor reduction due to the flea parasitic lifestyle. Based on these current data, Dr. Gillespie and colleagues posit that a dual mechanism of unequal crossing-over and gene conversion underpins flea genome variability (although the biological significance remains unknown). Further investigations of flea genome maintenance, regulation and evolution are on-going. The cat flea mitochondrial genome has no copy number variation, limiting this phenomenon to siphonapteran nuclear genomes.
Dr. Gillespie and colleagues also determined that cat fleas are often co-infected with divergent strains of Wolbachia (an intracellular bacterium related to Rickettsiae). From the generated C. felis sequence reads, closed genomes for two novel Wolbachia strains were assembled, annotated and analyzed. Both genomes contain laterally transferred genes that inform on the evolution of Wolbachia host associations. wCfeT carries biotin synthesis genes, while wCfeJ carries a CI-inducing toxin-antidote operon. Analyses of these genes highlight their mobility across the Wolbachia phylogeny and source to other intracellular bacteria. Additional screening of geographically diverse cat fleas revealed predominant co-infection (wCfeT/wCfeJ) amongst C. felis colonies, though single wCfeT infection is the norm in wild populations. Collectively, genomes of wCfeT and wCfeJ provide instances of lateral gene transfers that could drive transitions between parasitism and mutualism.
Awards and Affiliations
- 1998-2005: Member, Entomological Society of America
- 2000-present: Member, American Association for the Advancement of Science
- 2000-present: Member, Society for Molecular Biology and Evolution
- 2000-2003: Member, Society for the Study of Evolution
- 2000-present: Member, Society of Systematic Biologists
- 2004-2005: Member, European Society for Evolutionary Biology
- 2006-present: Member, American Society for Microbiology
- 2006-present: Member, American Society for Rickettsiology
Grants and Contracts
Vadyvaloo (PI), Eckstrand (Co-investigator), Gillespie (Co-investigator)
2/01/23 - 2/01/25
Emerging understanding of the rat flea response to Yersinia pestis infection
Gillespie (PI), Ernst (PI)
7/01/22 - 6/30/24
Investigating Rickettsia Interspecies and Host-Specific Lipopolysaccharide Variation
Gillespie (PI), Driscoll (PI)
1/22/21 - 12/31/22
Characterizing the Pan-genome of a Rickettsia Infecting the Eastern Black-legged Tick
Gillespie (PI), Rahman (PI)
5/23/19 - 4/30/21
Rickettsia cell envelope glycoconjugates are derived from the host cell amino sugar biosynthesis pathway
Gillespie (PI), Rahman (PI)
5/23/16 - 4/30/19
Characterizing gene family expansion in an atypical bacterial secretion system
- University of Maryland School of Medicine Council
- UMB Molecular Microbiology and Immunology Admissions Committee
- UMB Molecular Microbiology and Immunology Curriculum Committee
- Course master (with Dr. Ciaran Skerry) and Instructor for GPLS 710: Principles of Microbial Pathogenesis
- Course master (with Dr. Julie Hottop) and Instructor for GPLS 725: Advanced Microbial Pathogenesis
- Instructor for GPILS 693: Introduction to Molecular Microbiology and Immunology
- Instructor for RCR, CIPP907: Responsible Conduct of Research
- Judge for School of Medicine Annual Graduate Research Conference
- Judge for Microbiology and Immunology Graduate Research Competition
- Judge for School of Medicine Medical Student Research Day
Dr. Gillespie an ardent supporter of youth athletics, realizing the importance for introducing exercise and sports to children in a healthy and fun atmosphere. He coaches annually in various baseball, basketball and soccer programs in the Lutherville-Timonium area. He serves the community as the In-House Recreation Coordinator of the Lutherville-Timonium Soccer Club, and is a board member and commissioner in the Lutherville-Timonium Basketball Association. Dr. Gillespie is also a certified referee and judge for the Central Maryland Dive League.
- 2017-present: Editorial Board, PeerJ
- 2019-present: Reviewer, ZRG1 IDM-B 80 Study Section (Topics in Bacterial Pathogenesis), NIH
- 2021-present: Reviewer, ZRG1 IDIB-B 02 Study Section (Biology and Immunology of Bacteria and Other Pathogens), NIH
- 2018-present: Reviewer, Medical Research Council, United Kingdom Research and Innovation
- 2019: Site Visit Team Member for the review of Intramural Research Programs in the Tumor Vaccines and Biotechnology Branch (TVBB), Division of Cellular and Gene Therapies (DCGT), Office of Tissues and Advanced Therapies (OTAT), Center for Biologics Evaluation and Research (CBER), U.S. Food and Drug Administration, DHHS
- 2017-present: Scientific Review Committee, American Society for Rickettsiology
- 2020-present: Editorial Board, PLoS Neglected Tropical Diseases
- 2021-present: Editorial Board, Frontiers in Cellular and Infection Microbiology
- 2021-present: Reviewer, Maratona da Saúde (MdS), Infectious Diseases
- 2021-present: Reviewer, Bacterial Pathogenesis Study Section, NIH
- 2022-present: Reviewer, Prokaryotic Cell and Molecular Biology Study Section, NIH
Links of Interest
NCBI, The National Center for Biotechnology Information, advances science and health by providing access to biomedical and genomic information. NCBI's Pubmed is a repository for peer-reviewed science articles. Just search by key words or phrases; e.g., try searching with "anti-vaccine misinformation".
Bacterial and Viral Bioinformatics Resource Center (BV-BRC), an information system designed to support research on bacterial and viral infectious diseases. The BV-BRC combines the data and tools from the Legacy BRC resources: PATRIC, the bacterial BRC, and IRD and ViPR, the viral BRCs. Previously, PATRIC, the Pathosystems Resource Integration Center, provides integrated data and analysis tools to support biomedical research on bacterial infectious diseases. Dr. Gillespie was a team member and contributor to PATRIC from 2006 to 2013.
Dr. Jeanne Salje and her laboratories at University of Cambridge and Mahidol-Oxford Tropical Medicine Research Unit (MORU) in Bangkok, Thailand collaborate with Dr. Gillespie on projects analyzing rickettsial cell envelope biology, genomics, and evolution.
Dr. John Beckmann and his laboratory at Auburn University collaborate with Dr. Gillespie on several projects involving reproductive parasitism. Check out John's very inspiring art work and entertainment.
Dr. Bob Ernst, Dr. Alan Cross and Dr. Dave Goodlett collaborate with Dr. Gillespie on determining Rickettsia cell envelope glycoconjugate composition.
Dr. Greg Snyder (Institute of Human Virology) collaborates with Dr. Gillespie on protein structural biology.
Dr. Viveka Vadyvaloo and her laboratory at Washington State University collaborate with Dr. Gillespie on charcaterizing flea transcriptional responses to Yersinia and Rickettsia pathogens.
Dr. Anders Omsland and his laboratory at Washington State University collaborate with Dr. Gillespie on developing axenic culture media to support cell-free Rickettsia growth.
Dr. Tim Driscoll and his Vector-borne Pathogen Dynamics Laboratory at West Virginia University collaborate with Dr. Gillespie on many different projects, but especially those involving arthropod and bacteria genome sequencing, phylogenomics, and arthropod-bacteria interactions.
Luke J. Tallon, Scientific Director of the Genomics Resource Center, Institute for Genome Sciences, collaborates with Dr. Gillespie on the genome sequencing of the cat flea (Ctenocephalides felis) and its Wolbachia parasites.
Utilized Informatics Resources
Designing primers for PCR
Six frames DNA translation
Generate a CDS alignment from aligned proteins
Calculate positive selection
Protein sequence and functional information database
Searching for protein similarity
Build orthologous protein groups
Computing multiple sequence alignment
View and edit alignments
Manipulate/convert sequence formats
Masking multiple sequence alignments
Calculate pairwise divergences
Drawing phylogenetic trees
Repository for protein structures
Searching for structural analogs
Threading proteins to structures
Compare protein structures in 3D
AlphaFold Protein Structure Database
Predict SEC and lipoprotein signal sequences
Predict transmembrane spanning regions
Predict protein domains and other features
Making sequence logos
Draw gene/protein schemas
Visualize genomic data and information in a circular format
Bin and visualize metagenomic data
Tool for parasitology
Learn about the bacterial world
Dr. Gillespie Interviewed on Netflix Series
Dr. Gillespie shares the dangers posed by Chigoe Flea infestations in this video segment from the Netflix series, "72 Dangerous Animals: Latin America" (Season 2, Episode 7)
Dr. Gillespie and Dr. Mark Guillotte engage in advanced laboratory training techniques (filmed by Dr. Stephanie Lehman).
Dr. Gillespie and Dr. Verhoeve recently participated in the American Cancer Society-Diversity in Cancer Research Internship Program and trained Trinity Soto, a rising senior at Towson University (link to video)