From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma

Joseph Lykins; Mathew J. Moschitto; Ying Zhou; Ekaterina V. Filippova; Hoang V. Le; Tadakimi Tomita; Barbara A. Fox; David J. Bzik; Chunlei Su; Seesandra V. Rajagopala; Kristin Flores; Furio Spano; Stuart Woods; Craig W. Roberts; Cong Hua; Kamal El Bissati; Kelsey M. Wheeler; Sarah Dovgin; Stephen P. Muench; Martin McPhillie; Colin W.G. Fishwick; Wayne F.  Anderson; Patricia J. Lee; Mark Hickman; Louis M. Weiss; Jitender P. Dubey; Hernan A. Lorenzi; Richard B. Silverman; Rima L. McLeod

doi:10.1016/j.isci.2023.108477

From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma

Joseph Lykins
, Mathew J. Moschitto
, Ying Zhou
, Ekaterina V. Filippova
, Hoang V. Le
, Tadakimi Tomita
, Barbara A. Fox
, David J. Bzik
, Chunlei Su
, Seesandra V. Rajagopala
, Kristin Flores
, Furio Spano
, Stuart Woods
, Craig W. Roberts
, Cong Hua
, Kamal El Bissati
, Kelsey M. Wheeler
, Sarah Dovgin
, Stephen P. Muench
, Martin McPhillie

Colin W.G. Fishwick, Wayne F. Anderson, Patricia J. Lee, Mark Hickman, Louis M. Weiss, Jitender P. Dubey, Hernan A. Lorenzi, Richard B. Silverman, Rima L. McLeod

Strathclyde Institute Of Pharmacy And Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

25 Downloads (Pure)

Abstract

Toxoplasma gondii causes morbidity, mortality, and disseminates widely via cat sexual stages. Here, we find T. gondii ornithine aminotransferase (OAT) is conserved across phyla. We solve TgO/GABA-AT structures with bound inactivators at 1.55 Å and identify an inactivator selective for TgO/GABA-AT over human OAT and GABA-AT. However, abrogating TgO/GABA-AT genetically does not diminish replication, virulence, cyst-formation, or eliminate cat’s oocyst shedding. Increased sporozoite/merozoite TgO/GABA-AT expression led to our study of a mutagenized clone with oocyst formation blocked, arresting after forming male and female gametes, with “Rosetta stone”-like mutations in genes expressed in merozoites. Mutations are similar to those in organisms from plants to mammals, causing defects in conception and zygote formation, affecting merozoite capacitation, pH/ionicity/sodium-GABA concentrations, drawing attention to cyclic AMP/PKA, and genes enhancing energy or substrate formation in TgO/GABA-AT-related-pathways. These candidates potentially influence merozoite’s capacity to make gametes that fuse to become zygotes, thereby contaminating environments and causing disease.

Original language	English
Article number	108477
Number of pages	54
Journal	iScience
Volume	27
Issue number	1
Early online date	21 Dec 2023
DOIs	https://doi.org/10.1016/j.isci.2023.108477
Publication status	Published - 19 Jan 2024

Funding

The data collection was performed at the LS-CAT at the Advanced Photon Source supported by the Argonne National Laboratory operated by University of Chicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. The LS-CAT Sector 21 is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817 ). This project has been funded with Federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services , under Contracts No. HHSN272200700058C and HHSN272201200026C (W.F.A.). This research was supported, in part, by NIH NIDDK grant #1T35DK062719-27 (to J.L.) and by NIH grant R01 DA030604 (to R.B.S.). This work was also supported by the Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases (grant number U01AI082180-01,RMc ), the Conte Center, and the National Institutes of Health (grant numbers 1P50MH094267 and U01HL108634-01 ). This work also was supported by NIH grant AI155603 (DJB) and NIH grant AI172811 (D.J.B.). We thank the Mann Cornwell family, Engel family (and “Taking Out Toxo”), Morel, Pritzker, Allen, Drago, VanDusen/Longfellow, Rodriguez and Rooney families for their support of this work. We thank Andrew Grose for assistance with references during manuscript preparation. We thank Michael Grigg for helpful discussion. We thank L. Knoll and B. Martorelli Di Genova for their early work with their model and image in Figure 7 A. We thank Mario Falchi (Core Facilities, ISS) for confocal microscope analysis. We thank Mohamed-Ali Hakimi for advice concerning the MORC analysis and permission to include information from the data in that analysis. The data collection was performed at the LS-CAT at the Advanced Photon Source supported by the Argonne National Laboratory operated by University of Chicago Argonne, LLC, for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. The LS-CAT Sector 21 is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). This project has been funded with Federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under Contracts No. HHSN272200700058C and HHSN272201200026C (W.F.A.). This research was supported, in part, by NIH NIDDK grant #1T35DK062719-27 (to J.L.) and by NIH grant R01 DA030604 (to R.B.S.). This work was also supported by the Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases (grant number U01AI082180-01,RMc), the Conte Center, and the National Institutes of Health (grant numbers 1P50MH094267 and U01HL108634-01). This work also was supported by NIH grant AI155603 (DJB) and NIH grant AI172811 (D.J.B.). We thank the Mann Cornwell family, Engel family (and “Taking Out Toxo”), Morel, Pritzker, Allen, Drago, VanDusen/Longfellow, Rodriguez and Rooney families for their support of this work. We thank Andrew Grose for assistance with references during manuscript preparation. We thank Michael Grigg for helpful discussion. We thank L. Knoll and B. Martorelli Di Genova for their early work with their model and image in Figure 7A. We thank Mario Falchi (Core Facilities, ISS) for confocal microscope analysis. We thank Mohamed-Ali Hakimi for advice concerning the MORC analysis and permission to include information from the data in that analysis. Conception and design: R.L.M. R.B.S. H.A.L. L.M.W. D.B. J.L. J.P.D. E.V.F. T.T. B.A.F. D.B. and C.S. Performed and/or interpreted experiments: Y.Z. M.J.M. H.V.L. E.V.F. W.A. S.D. H.A.L. S.W. C.W.R. P.L. M.H. B.F. K.E.B. F.S. C.H. P.L. M.H. M.M. J.P.D. M.M. S.V.R. K.F. K.E.B. K.M.W. S.D. C.H. S.P.M. M.M. and R.B.S. Wrote manuscript: R.L.M. M.J.M. H.V.L. E.V.F. J.L. D.B. L.W. H.A.L. C.W.R. and R.B.S. Revised manuscript: R.B.S. M.J.M. R.L.M. E.V.F. H.V.L. E.V.F. H.A.L. J.L. Y.Z. J.P.D. and C.W.R. H.V.L. is currently a program officer at the National Institute on Drug Abuse at the National Institutes of Health. H.A.L. is currently a scientist working at National Institute of Diabetes, Digestive and Kidney Disease (NIDDK) at the National Institutes of Health. The findings and conclusions of this article are those of the authors and do not necessarily reflect the views of the National Institute on Drug Abuse, National Institute of Diabetes, Digestive and Kidney Diseases (NIDDK), the National Institutes of Health, nor the US Department of Health and Human Services. R.B.S. and R.L.M. with H.V.L. submitted Patent Number: US 10,632,088 B2 INACTIVATORS OF TOXOPLASMA GONDII ORNITHINE AMINOTRANSFERASE FOR TREATING TOXOPLASMOSIS AND MALARIA through their Technology Transfer offices. The authors declare no competing financial interest. We support inclusion, diverse and equitable conduct of research.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

parasitology
cell biology
Toxoplasma gondii
toxoplasmosis
ornithine aminotransferase
gametes

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10.1016/j.isci.2023.108477Licence: CC BY-NC-ND 4.0

Lykins-etal-iScience-2023-From-TgO-GABA-AT-GABA-and-T-263-mutant-to-conception-of-ToxoplasmaFinal published version, 13.5 MBLicence: CC BY-NC-ND 4.0

Cite this

Lykins, J., Moschitto, M. J., Zhou, Y., Filippova, E. V., Le, H. V., Tomita, T., Fox, B. A., Bzik, D. J., Su, C., Rajagopala, S. V., Flores, K., Spano, F., Woods, S., Roberts, C. W., Hua, C., El Bissati, K., Wheeler, K. M., Dovgin, S., Muench, S. P., ... McLeod, R. L. (2024). From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma. iScience, 27(1), Article 108477. https://doi.org/10.1016/j.isci.2023.108477

@article{15adb73bb92047429789e4a438a6dc8d,

title = "From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma",

abstract = "Toxoplasma gondii causes morbidity, mortality, and disseminates widely via cat sexual stages. Here, we find T. gondii ornithine aminotransferase (OAT) is conserved across phyla. We solve TgO/GABA-AT structures with bound inactivators at 1.55 {\AA} and identify an inactivator selective for TgO/GABA-AT over human OAT and GABA-AT. However, abrogating TgO/GABA-AT genetically does not diminish replication, virulence, cyst-formation, or eliminate cat{\textquoteright}s oocyst shedding. Increased sporozoite/merozoite TgO/GABA-AT expression led to our study of a mutagenized clone with oocyst formation blocked, arresting after forming male and female gametes, with “Rosetta stone”-like mutations in genes expressed in merozoites. Mutations are similar to those in organisms from plants to mammals, causing defects in conception and zygote formation, affecting merozoite capacitation, pH/ionicity/sodium-GABA concentrations, drawing attention to cyclic AMP/PKA, and genes enhancing energy or substrate formation in TgO/GABA-AT-related-pathways. These candidates potentially influence merozoite{\textquoteright}s capacity to make gametes that fuse to become zygotes, thereby contaminating environments and causing disease.",

keywords = "parasitology, cell biology, Toxoplasma gondii, toxoplasmosis, ornithine aminotransferase, gametes",

author = "Joseph Lykins and Moschitto, \{Mathew J.\} and Ying Zhou and Filippova, \{Ekaterina V.\} and Le, \{Hoang V.\} and Tadakimi Tomita and Fox, \{Barbara A.\} and Bzik, \{David J.\} and Chunlei Su and Rajagopala, \{Seesandra V.\} and Kristin Flores and Furio Spano and Stuart Woods and Roberts, \{Craig W.\} and Cong Hua and \{El Bissati\}, Kamal and Wheeler, \{Kelsey M.\} and Sarah Dovgin and Muench, \{Stephen P.\} and Martin McPhillie and Fishwick, \{Colin W.G.\} and Anderson, \{Wayne F.\} and Lee, \{Patricia J.\} and Mark Hickman and Weiss, \{Louis M.\} and Dubey, \{Jitender P.\} and Lorenzi, \{Hernan A.\} and Silverman, \{Richard B.\} and McLeod, \{Rima L.\}",

year = "2024",

month = jan,

day = "19",

doi = "10.1016/j.isci.2023.108477",

language = "English",

volume = "27",

journal = "iScience",

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Lykins, J, Moschitto, MJ, Zhou, Y, Filippova, EV, Le, HV, Tomita, T, Fox, BA, Bzik, DJ, Su, C, Rajagopala, SV, Flores, K, Spano, F, Woods, S, Roberts, CW, Hua, C, El Bissati, K, Wheeler, KM, Dovgin, S, Muench, SP, McPhillie, M, Fishwick, CWG, Anderson, WF, Lee, PJ, Hickman, M, Weiss, LM, Dubey, JP, Lorenzi, HA, Silverman, RB & McLeod, RL 2024, 'From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma', iScience, vol. 27, no. 1, 108477. https://doi.org/10.1016/j.isci.2023.108477

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T1 - From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma

AU - Lykins, Joseph

AU - Moschitto, Mathew J.

AU - Zhou, Ying

AU - Filippova, Ekaterina V.

AU - Le, Hoang V.

AU - Tomita, Tadakimi

AU - Fox, Barbara A.

AU - Bzik, David J.

AU - Su, Chunlei

AU - Rajagopala, Seesandra V.

AU - Flores, Kristin

AU - Spano, Furio

AU - Woods, Stuart

AU - Roberts, Craig W.

AU - Hua, Cong

AU - El Bissati, Kamal

AU - Wheeler, Kelsey M.

AU - Dovgin, Sarah

AU - Muench, Stephen P.

AU - McPhillie, Martin

AU - Fishwick, Colin W.G.

AU - Anderson, Wayne F.

AU - Lee, Patricia J.

AU - Hickman, Mark

AU - Weiss, Louis M.

AU - Dubey, Jitender P.

AU - Lorenzi, Hernan A.

AU - Silverman, Richard B.

AU - McLeod, Rima L.

PY - 2024/1/19

Y1 - 2024/1/19

N2 - Toxoplasma gondii causes morbidity, mortality, and disseminates widely via cat sexual stages. Here, we find T. gondii ornithine aminotransferase (OAT) is conserved across phyla. We solve TgO/GABA-AT structures with bound inactivators at 1.55 Å and identify an inactivator selective for TgO/GABA-AT over human OAT and GABA-AT. However, abrogating TgO/GABA-AT genetically does not diminish replication, virulence, cyst-formation, or eliminate cat’s oocyst shedding. Increased sporozoite/merozoite TgO/GABA-AT expression led to our study of a mutagenized clone with oocyst formation blocked, arresting after forming male and female gametes, with “Rosetta stone”-like mutations in genes expressed in merozoites. Mutations are similar to those in organisms from plants to mammals, causing defects in conception and zygote formation, affecting merozoite capacitation, pH/ionicity/sodium-GABA concentrations, drawing attention to cyclic AMP/PKA, and genes enhancing energy or substrate formation in TgO/GABA-AT-related-pathways. These candidates potentially influence merozoite’s capacity to make gametes that fuse to become zygotes, thereby contaminating environments and causing disease.

AB - Toxoplasma gondii causes morbidity, mortality, and disseminates widely via cat sexual stages. Here, we find T. gondii ornithine aminotransferase (OAT) is conserved across phyla. We solve TgO/GABA-AT structures with bound inactivators at 1.55 Å and identify an inactivator selective for TgO/GABA-AT over human OAT and GABA-AT. However, abrogating TgO/GABA-AT genetically does not diminish replication, virulence, cyst-formation, or eliminate cat’s oocyst shedding. Increased sporozoite/merozoite TgO/GABA-AT expression led to our study of a mutagenized clone with oocyst formation blocked, arresting after forming male and female gametes, with “Rosetta stone”-like mutations in genes expressed in merozoites. Mutations are similar to those in organisms from plants to mammals, causing defects in conception and zygote formation, affecting merozoite capacitation, pH/ionicity/sodium-GABA concentrations, drawing attention to cyclic AMP/PKA, and genes enhancing energy or substrate formation in TgO/GABA-AT-related-pathways. These candidates potentially influence merozoite’s capacity to make gametes that fuse to become zygotes, thereby contaminating environments and causing disease.

KW - parasitology

KW - cell biology

KW - Toxoplasma gondii

KW - toxoplasmosis

KW - ornithine aminotransferase

KW - gametes

U2 - 10.1016/j.isci.2023.108477

DO - 10.1016/j.isci.2023.108477

M3 - Article

SN - 2589-0042

VL - 27

JO - iScience

JF - iScience

IS - 1

M1 - 108477

ER -

From TgO/GABA-AT, GABA, and T-263 mutant to conception of Toxoplasma

Abstract

Funding

UN SDGs

Keywords

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