Evolutionary conservation and in vitro reconstitution of microsporidian iron-sulfur cluster biosynthesis

Sven-A Freibert; Alina V. Goldberg; Christian Hacker; Sabine  Molik; Paul Dean; Tom A. Williams; Sirintra Nakjang; Shaojun Long; Kacper Sendra; Eckhard Bill; Eva Heinz; Robert P. Hirt; John Milton Lucocq; T. Martin Embley; Roland Lill

doi:10.1038/ncomms13932

Evolutionary conservation and in vitro reconstitution of microsporidian iron-sulfur cluster biosynthesis

Sven-A Freibert, Alina V. Goldberg, Christian Hacker, Sabine Molik, Paul Dean, Tom A. Williams, Sirintra Nakjang, Shaojun Long, Kacper Sendra, Eckhard Bill, Eva Heinz, Robert P. Hirt, John Milton Lucocq, T. Martin Embley, Roland Lill

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Abstract

Microsporidians are a diverse group of obligate intracellular parasites that have minimized their genome content and simplified their sub-cellular structures by reductive evolution. Functional studies are limited because we lack reliable genetic tools for their manipulation. Here, we demonstrate that the cristae-deficient mitochondrion (mitosome) of the microsporidian Trachipleistophora hominis is the functional site of iron-sulphur cluster (ISC) assembly, which we suggest is the essential task of this organelle. Cell fractionation, fluorescence imaging and fine-scale immunoelectron microscopy demonstrate that mitosomes contain a complete pathway for [2Fe-2S] cluster biosynthesis that we biochemically reconstituted using purified recombinant mitosomal ISC proteins. Reconstitution proceeded as rapidly and efficiently as observed for yeast or fungal mitochondrial ISC components. Core components of the T. hominis cytosolic iron-sulphur protein assembly (CIA) pathway were also identified including the essential Cfd1-Nbp35 scaffold complex that assembles a [4Fe-4S] cluster as shown by spectroscopic methods in vitro. Phylogenetic analyses reveal that both the ISC and CIA biosynthetic pathways are predominantly bacterial, but their cytosolic and nuclear target Fe/S proteins are mainly archaeal. This mixed evolutionary history of the Fe/S-related proteins and pathways, and their strong conservation among highly reduced parasites, provides additional compelling evidence for the ancient chimeric ancestry of eukaryotes.

Original language	English
Article number	13932
Number of pages	12
Journal	Nature Communications
Volume	8
DOIs	https://doi.org/10.1038/ncomms13932
Publication status	Published - 4 Jan 2017

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Freibert, S.-A., Goldberg, A. V., Hacker, C., Molik, S., Dean, P., Williams, T. A., Nakjang, S., Long, S., Sendra, K., Bill, E., Heinz, E., Hirt, R. P., Lucocq, J. M., Embley, T. M., & Lill, R. (2017). Evolutionary conservation and in vitro reconstitution of microsporidian iron-sulfur cluster biosynthesis. Nature Communications, 8, Article 13932. https://doi.org/10.1038/ncomms13932

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title = "Evolutionary conservation and in vitro reconstitution of microsporidian iron-sulfur cluster biosynthesis",

abstract = "Microsporidians are a diverse group of obligate intracellular parasites that have minimized their genome content and simplified their sub-cellular structures by reductive evolution. Functional studies are limited because we lack reliable genetic tools for their manipulation. Here, we demonstrate that the cristae-deficient mitochondrion (mitosome) of the microsporidian Trachipleistophora hominis is the functional site of iron-sulphur cluster (ISC) assembly, which we suggest is the essential task of this organelle. Cell fractionation, fluorescence imaging and fine-scale immunoelectron microscopy demonstrate that mitosomes contain a complete pathway for [2Fe-2S] cluster biosynthesis that we biochemically reconstituted using purified recombinant mitosomal ISC proteins. Reconstitution proceeded as rapidly and efficiently as observed for yeast or fungal mitochondrial ISC components. Core components of the T. hominis cytosolic iron-sulphur protein assembly (CIA) pathway were also identified including the essential Cfd1-Nbp35 scaffold complex that assembles a [4Fe-4S] cluster as shown by spectroscopic methods in vitro. Phylogenetic analyses reveal that both the ISC and CIA biosynthetic pathways are predominantly bacterial, but their cytosolic and nuclear target Fe/S proteins are mainly archaeal. This mixed evolutionary history of the Fe/S-related proteins and pathways, and their strong conservation among highly reduced parasites, provides additional compelling evidence for the ancient chimeric ancestry of eukaryotes.",

author = "Sven-A Freibert and Goldberg, {Alina V.} and Christian Hacker and Sabine Molik and Paul Dean and Williams, {Tom A.} and Sirintra Nakjang and Shaojun Long and Kacper Sendra and Eckhard Bill and Eva Heinz and Hirt, {Robert P.} and Lucocq, {John Milton} and Embley, {T. Martin} and Roland Lill",

note = "This work was supported by Marie Curie Postdoctoral Fellowships to T.A.W., E. H. and S. L., a European Research Council Advanced Investigator Grant (ERC-2010-AdG-268701) to T.M.E., and a Wellcome Trust Programme Grant (number 045404) to T.M.E. and J.M.L. R.L. acknowledges generous financial support from Deutsche Forschungsgemeinschaft (SFB 593, SFB 987, GRK 1216, LI 415/5), LOEWE program of state Hessen, Max-Planck Gesellschaft, von Behring-R{\"o}ntgen Stiftung",

year = "2017",

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doi = "10.1038/ncomms13932",

language = "English",

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T1 - Evolutionary conservation and in vitro reconstitution of microsporidian iron-sulfur cluster biosynthesis

AU - Freibert, Sven-A

AU - Goldberg, Alina V.

AU - Hacker, Christian

AU - Molik, Sabine

AU - Dean, Paul

AU - Williams, Tom A.

AU - Nakjang, Sirintra

AU - Long, Shaojun

AU - Sendra, Kacper

AU - Bill, Eckhard

AU - Heinz, Eva

AU - Hirt, Robert P.

AU - Lucocq, John Milton

AU - Embley, T. Martin

AU - Lill, Roland

N1 - This work was supported by Marie Curie Postdoctoral Fellowships to T.A.W., E. H. and S. L., a European Research Council Advanced Investigator Grant (ERC-2010-AdG-268701) to T.M.E., and a Wellcome Trust Programme Grant (number 045404) to T.M.E. and J.M.L. R.L. acknowledges generous financial support from Deutsche Forschungsgemeinschaft (SFB 593, SFB 987, GRK 1216, LI 415/5), LOEWE program of state Hessen, Max-Planck Gesellschaft, von Behring-Röntgen Stiftung

PY - 2017/1/4

Y1 - 2017/1/4

N2 - Microsporidians are a diverse group of obligate intracellular parasites that have minimized their genome content and simplified their sub-cellular structures by reductive evolution. Functional studies are limited because we lack reliable genetic tools for their manipulation. Here, we demonstrate that the cristae-deficient mitochondrion (mitosome) of the microsporidian Trachipleistophora hominis is the functional site of iron-sulphur cluster (ISC) assembly, which we suggest is the essential task of this organelle. Cell fractionation, fluorescence imaging and fine-scale immunoelectron microscopy demonstrate that mitosomes contain a complete pathway for [2Fe-2S] cluster biosynthesis that we biochemically reconstituted using purified recombinant mitosomal ISC proteins. Reconstitution proceeded as rapidly and efficiently as observed for yeast or fungal mitochondrial ISC components. Core components of the T. hominis cytosolic iron-sulphur protein assembly (CIA) pathway were also identified including the essential Cfd1-Nbp35 scaffold complex that assembles a [4Fe-4S] cluster as shown by spectroscopic methods in vitro. Phylogenetic analyses reveal that both the ISC and CIA biosynthetic pathways are predominantly bacterial, but their cytosolic and nuclear target Fe/S proteins are mainly archaeal. This mixed evolutionary history of the Fe/S-related proteins and pathways, and their strong conservation among highly reduced parasites, provides additional compelling evidence for the ancient chimeric ancestry of eukaryotes.

AB - Microsporidians are a diverse group of obligate intracellular parasites that have minimized their genome content and simplified their sub-cellular structures by reductive evolution. Functional studies are limited because we lack reliable genetic tools for their manipulation. Here, we demonstrate that the cristae-deficient mitochondrion (mitosome) of the microsporidian Trachipleistophora hominis is the functional site of iron-sulphur cluster (ISC) assembly, which we suggest is the essential task of this organelle. Cell fractionation, fluorescence imaging and fine-scale immunoelectron microscopy demonstrate that mitosomes contain a complete pathway for [2Fe-2S] cluster biosynthesis that we biochemically reconstituted using purified recombinant mitosomal ISC proteins. Reconstitution proceeded as rapidly and efficiently as observed for yeast or fungal mitochondrial ISC components. Core components of the T. hominis cytosolic iron-sulphur protein assembly (CIA) pathway were also identified including the essential Cfd1-Nbp35 scaffold complex that assembles a [4Fe-4S] cluster as shown by spectroscopic methods in vitro. Phylogenetic analyses reveal that both the ISC and CIA biosynthetic pathways are predominantly bacterial, but their cytosolic and nuclear target Fe/S proteins are mainly archaeal. This mixed evolutionary history of the Fe/S-related proteins and pathways, and their strong conservation among highly reduced parasites, provides additional compelling evidence for the ancient chimeric ancestry of eukaryotes.

U2 - 10.1038/ncomms13932

DO - 10.1038/ncomms13932

M3 - Article

SN - 2041-1723

VL - 8

JO - Nature Communications

JF - Nature Communications

M1 - 13932

ER -

Evolutionary conservation and in vitro reconstitution of microsporidian iron-sulfur cluster biosynthesis

Abstract

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