Cryo-EM data processing workflow.
<div><p>[Fe-S] clusters are ancient and ubiquitous protein co-factors, which contributed to the emergence of life in an anoxic planet. We have recently identified two minimal [Fe-S] biogenesis systems, MIS and SMS, inferred to be ancestral systems dating back to the Last Universal Common...
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2025
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| _version_ | 1852019037843750912 |
|---|---|
| author | Macha Dussouchaud (21601445) |
| author2 | Markel Martinez-Carranza (21601448) Pierre-Simon Garcia (21601451) Martin Clémancey (347878) Geneviève Blondin (2668756) Jean Michel Betton (21601454) Ahmed Haouz (5624486) Simonetta Gribaldo (3252) Sandrine Ollagnier de Choudens (21601457) Ludovic Sauguet (250490) Ariel Mechaly (12131742) Frédéric Barras (207624) |
| author2_role | author author author author author author author author author author author |
| author_facet | Macha Dussouchaud (21601445) Markel Martinez-Carranza (21601448) Pierre-Simon Garcia (21601451) Martin Clémancey (347878) Geneviève Blondin (2668756) Jean Michel Betton (21601454) Ahmed Haouz (5624486) Simonetta Gribaldo (3252) Sandrine Ollagnier de Choudens (21601457) Ludovic Sauguet (250490) Ariel Mechaly (12131742) Frédéric Barras (207624) |
| author_role | author |
| dc.creator.none.fl_str_mv | Macha Dussouchaud (21601445) Markel Martinez-Carranza (21601448) Pierre-Simon Garcia (21601451) Martin Clémancey (347878) Geneviève Blondin (2668756) Jean Michel Betton (21601454) Ahmed Haouz (5624486) Simonetta Gribaldo (3252) Sandrine Ollagnier de Choudens (21601457) Ludovic Sauguet (250490) Ariel Mechaly (12131742) Frédéric Barras (207624) |
| dc.date.none.fl_str_mv | 2025-06-25T17:46:41Z |
| dc.identifier.none.fl_str_mv | 10.1371/journal.pbio.3003223.s005 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/figure/Cryo-EM_data_processing_workflow_/29405915 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biophysics Biochemistry Microbiology Genetics Biotechnology Evolutionary Biology Cancer Infectious Diseases Virology Chemical Sciences not elsewhere classified ubiquitous protein co studied modern iron present study focuses mutually exclusive allowing methanocaldococcus jannaschii </ mechanistic model describing escherichia coli </ contained cys residues bona fide </ anaerobic prokaryotes prior structural studies showed study established sms sms system rescued smsc subunit hosts suf systems grown cluster binding sites cluster assembly site biogenesis system sms biogenesis system biogenesis systems unique structural cluster assembly ancestor sms sulfur cluster unique mechanism similar c regulatory coupling nif ), key role hyperthermophilic archaeon heterotetratmer wherein gave rise functional features anoxic planet anoxic conditions |
| dc.title.none.fl_str_mv | Cryo-EM data processing workflow. |
| dc.type.none.fl_str_mv | Image Figure info:eu-repo/semantics/publishedVersion image |
| description | <div><p>[Fe-S] clusters are ancient and ubiquitous protein co-factors, which contributed to the emergence of life in an anoxic planet. We have recently identified two minimal [Fe-S] biogenesis systems, MIS and SMS, inferred to be ancestral systems dating back to the Last Universal Common Ancestor and which gave rise to the well-studied modern Iron-Sulfur Cluster (ISC), Nitrogen Fixation (NIF), and Sulfur Mobilization (SUF) machineries. The present study focuses on the ancestor SMS from the hyperthermophilic archaeon <i>Methanocaldococcus jannaschii</i>. Biochemical and structural studies showed that SMS is made of a SmsC<sub>2</sub>B<sub>2</sub> heterotetratmer wherein the SmsC subunit hosts both ATP and [Fe-S] cluster binding sites. Binding of ATP and assembly of [Fe-S] were found to be mutually exclusive allowing for a regulatory coupling between binding of both substrates. Mutagenesis and in vitro transfer experiments revealed the key role of SmsC-contained Cys residues in cluster assembly. Strikingly, the SMS system rescued a non-viable <i>Escherichia coli</i> strain lacking endogenous ISC and SUF systems grown under anoxic conditions, in the presence of Na<sub>2</sub>S, indicating that sulfide is a source of sulfur for SMS. In addition, we predict that most archaea SmsC proteins hold a similar C-terminal [Fe-S] cluster assembly site. Taking into account those unique structural and functional features, we propose a mechanistic model describing how SmsC<sub>2</sub>B<sub>2</sub> assembles and distributes [4Fe-4S] clusters. Altogether this study established SMS as a new <i>bona fide</i> [Fe-S] biogenesis system that operated in anaerobic prokaryotes prior to evolve to SUF after the Great Oxydation Event.</p></div> |
| eu_rights_str_mv | openAccess |
| id | Manara_d4ebd63bbcd2b8a6bc94d4b226bced1f |
| identifier_str_mv | 10.1371/journal.pbio.3003223.s005 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/29405915 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Cryo-EM data processing workflow.Macha Dussouchaud (21601445)Markel Martinez-Carranza (21601448)Pierre-Simon Garcia (21601451)Martin Clémancey (347878)Geneviève Blondin (2668756)Jean Michel Betton (21601454)Ahmed Haouz (5624486)Simonetta Gribaldo (3252)Sandrine Ollagnier de Choudens (21601457)Ludovic Sauguet (250490)Ariel Mechaly (12131742)Frédéric Barras (207624)BiophysicsBiochemistryMicrobiologyGeneticsBiotechnologyEvolutionary BiologyCancerInfectious DiseasesVirologyChemical Sciences not elsewhere classifiedubiquitous protein costudied modern ironpresent study focusesmutually exclusive allowingmethanocaldococcus jannaschii </mechanistic model describingescherichia coli </contained cys residuesbona fide </anaerobic prokaryotes priorstructural studies showedstudy established smssms system rescuedsmsc subunit hostssuf systems growncluster binding sitescluster assembly sitebiogenesis system smsbiogenesis systembiogenesis systemsunique structuralcluster assemblyancestor smssulfur clusterunique mechanismsimilar cregulatory couplingnif ),key rolehyperthermophilic archaeonheterotetratmer whereingave risefunctional featuresanoxic planetanoxic conditions<div><p>[Fe-S] clusters are ancient and ubiquitous protein co-factors, which contributed to the emergence of life in an anoxic planet. We have recently identified two minimal [Fe-S] biogenesis systems, MIS and SMS, inferred to be ancestral systems dating back to the Last Universal Common Ancestor and which gave rise to the well-studied modern Iron-Sulfur Cluster (ISC), Nitrogen Fixation (NIF), and Sulfur Mobilization (SUF) machineries. The present study focuses on the ancestor SMS from the hyperthermophilic archaeon <i>Methanocaldococcus jannaschii</i>. Biochemical and structural studies showed that SMS is made of a SmsC<sub>2</sub>B<sub>2</sub> heterotetratmer wherein the SmsC subunit hosts both ATP and [Fe-S] cluster binding sites. Binding of ATP and assembly of [Fe-S] were found to be mutually exclusive allowing for a regulatory coupling between binding of both substrates. Mutagenesis and in vitro transfer experiments revealed the key role of SmsC-contained Cys residues in cluster assembly. Strikingly, the SMS system rescued a non-viable <i>Escherichia coli</i> strain lacking endogenous ISC and SUF systems grown under anoxic conditions, in the presence of Na<sub>2</sub>S, indicating that sulfide is a source of sulfur for SMS. In addition, we predict that most archaea SmsC proteins hold a similar C-terminal [Fe-S] cluster assembly site. Taking into account those unique structural and functional features, we propose a mechanistic model describing how SmsC<sub>2</sub>B<sub>2</sub> assembles and distributes [4Fe-4S] clusters. Altogether this study established SMS as a new <i>bona fide</i> [Fe-S] biogenesis system that operated in anaerobic prokaryotes prior to evolve to SUF after the Great Oxydation Event.</p></div>2025-06-25T17:46:41ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pbio.3003223.s005https://figshare.com/articles/figure/Cryo-EM_data_processing_workflow_/29405915CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/294059152025-06-25T17:46:41Z |
| spellingShingle | Cryo-EM data processing workflow. Macha Dussouchaud (21601445) Biophysics Biochemistry Microbiology Genetics Biotechnology Evolutionary Biology Cancer Infectious Diseases Virology Chemical Sciences not elsewhere classified ubiquitous protein co studied modern iron present study focuses mutually exclusive allowing methanocaldococcus jannaschii </ mechanistic model describing escherichia coli </ contained cys residues bona fide </ anaerobic prokaryotes prior structural studies showed study established sms sms system rescued smsc subunit hosts suf systems grown cluster binding sites cluster assembly site biogenesis system sms biogenesis system biogenesis systems unique structural cluster assembly ancestor sms sulfur cluster unique mechanism similar c regulatory coupling nif ), key role hyperthermophilic archaeon heterotetratmer wherein gave rise functional features anoxic planet anoxic conditions |
| status_str | publishedVersion |
| title | Cryo-EM data processing workflow. |
| title_full | Cryo-EM data processing workflow. |
| title_fullStr | Cryo-EM data processing workflow. |
| title_full_unstemmed | Cryo-EM data processing workflow. |
| title_short | Cryo-EM data processing workflow. |
| title_sort | Cryo-EM data processing workflow. |
| topic | Biophysics Biochemistry Microbiology Genetics Biotechnology Evolutionary Biology Cancer Infectious Diseases Virology Chemical Sciences not elsewhere classified ubiquitous protein co studied modern iron present study focuses mutually exclusive allowing methanocaldococcus jannaschii </ mechanistic model describing escherichia coli </ contained cys residues bona fide </ anaerobic prokaryotes prior structural studies showed study established sms sms system rescued smsc subunit hosts suf systems grown cluster binding sites cluster assembly site biogenesis system sms biogenesis system biogenesis systems unique structural cluster assembly ancestor sms sulfur cluster unique mechanism similar c regulatory coupling nif ), key role hyperthermophilic archaeon heterotetratmer wherein gave rise functional features anoxic planet anoxic conditions |