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Panorama of ancient metazoan macromolecular complexes

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ABSTRACT

Macromolecular complexes are essential to conserved biological processes, but their prevalence across animals is unclear. By combining extensive biochemical fractionation with quantitative mass spectrometry, we directly examined the composition of soluble multiprotein complexes among diverse metazoan models. Using an integrative approach, we then generated a draft conservation map consisting of >1 million putative high-confidence co-complex interactions for species with fully sequenced genomes that encompasses functional modules present broadly across all extant animals. Clustering revealed a spectrum of conservation, ranging from ancient Eukaryal assemblies likely serving cellular housekeeping roles for at least 1 billion years, ancestral complexes that have accrued contemporary components, and rarer metazoan innovations linked to multicellularity. We validated these projections by independent co-fractionation experiments in evolutionarily distant species, by affinity-purification and by functional analyses. The comprehensiveness, centrality and modularity of these reconstructed interactomes reflect their fundamental mechanistic significance and adaptive value to animal cell systems.

No MeSH data available.


Related in: MedlinePlus

Supporting data for BUB3 and CCDC97 experimentsa, Sequence alignment showing conservation of ZNF207 GLEBS domain. b, Targeted CRISPR/Cas9 induced knockout of CCDC97 in two independent lines of human HEK293 cells, as verified by Western blotting (expanded gel images provided in SI), also results in a slight decrease in annotated SF3B3 component levels. c, Loss of CCDC97 impairs cell growth. Lines show growth curves of control versus knockout cell lines in two biological replicate assays.
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Figure 15: Supporting data for BUB3 and CCDC97 experimentsa, Sequence alignment showing conservation of ZNF207 GLEBS domain. b, Targeted CRISPR/Cas9 induced knockout of CCDC97 in two independent lines of human HEK293 cells, as verified by Western blotting (expanded gel images provided in SI), also results in a slight decrease in annotated SF3B3 component levels. c, Loss of CCDC97 impairs cell growth. Lines show growth curves of control versus knockout cell lines in two biological replicate assays.

Mentions: Among metazoan-specific protein complexes, we confirmed physical and functional associations of spindle checkpoint protein BUB3 with ZNF207, a zinc finger protein conspicuously lacking orthologs in cnidarians and fungi. ZNF207 binds Bub3 via a Gle2-binding-sequence (GLEBS) motif35 restricted to deuterostomes and protostomes (Extended Data Fig. 10a). As in human, knockdown of ZNF207 in C. elegans enhanced lethality due to impaired Bub3-mediated checkpoint arrest (Fig. 5c).


Panorama of ancient metazoan macromolecular complexes
Supporting data for BUB3 and CCDC97 experimentsa, Sequence alignment showing conservation of ZNF207 GLEBS domain. b, Targeted CRISPR/Cas9 induced knockout of CCDC97 in two independent lines of human HEK293 cells, as verified by Western blotting (expanded gel images provided in SI), also results in a slight decrease in annotated SF3B3 component levels. c, Loss of CCDC97 impairs cell growth. Lines show growth curves of control versus knockout cell lines in two biological replicate assays.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5036527&req=5

Figure 15: Supporting data for BUB3 and CCDC97 experimentsa, Sequence alignment showing conservation of ZNF207 GLEBS domain. b, Targeted CRISPR/Cas9 induced knockout of CCDC97 in two independent lines of human HEK293 cells, as verified by Western blotting (expanded gel images provided in SI), also results in a slight decrease in annotated SF3B3 component levels. c, Loss of CCDC97 impairs cell growth. Lines show growth curves of control versus knockout cell lines in two biological replicate assays.
Mentions: Among metazoan-specific protein complexes, we confirmed physical and functional associations of spindle checkpoint protein BUB3 with ZNF207, a zinc finger protein conspicuously lacking orthologs in cnidarians and fungi. ZNF207 binds Bub3 via a Gle2-binding-sequence (GLEBS) motif35 restricted to deuterostomes and protostomes (Extended Data Fig. 10a). As in human, knockdown of ZNF207 in C. elegans enhanced lethality due to impaired Bub3-mediated checkpoint arrest (Fig. 5c).

View Article: PubMed Central - PubMed

ABSTRACT

Macromolecular complexes are essential to conserved biological processes, but their prevalence across animals is unclear. By combining extensive biochemical fractionation with quantitative mass spectrometry, we directly examined the composition of soluble multiprotein complexes among diverse metazoan models. Using an integrative approach, we then generated a draft conservation map consisting of >1 million putative high-confidence co-complex interactions for species with fully sequenced genomes that encompasses functional modules present broadly across all extant animals. Clustering revealed a spectrum of conservation, ranging from ancient Eukaryal assemblies likely serving cellular housekeeping roles for at least 1 billion years, ancestral complexes that have accrued contemporary components, and rarer metazoan innovations linked to multicellularity. We validated these projections by independent co-fractionation experiments in evolutionarily distant species, by affinity-purification and by functional analyses. The comprehensiveness, centrality and modularity of these reconstructed interactomes reflect their fundamental mechanistic significance and adaptive value to animal cell systems.

No MeSH data available.


Related in: MedlinePlus